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1. Cell next page 13 Disassembling the XCell4 SureLock Midi Cell Procedure 14 Be sure the cables are disconnected from the power supply 1 2 Remove the lid Discard the buffer Unlock the XCell4 SureLock Assembly by moving the tension lever to the TM unlocked position indicated on the XCell4 SureLock Assembly Note Moving the tension lever to the unlocked position before discarding the buffer from the midi cell may cause some splashing of the buffer Prior to unlocking the XCell4 SureLock Assembly be sure to discard the buffer Remove the Buffer Cores with the gel cassettes from the Lower Buffer Chamber while holding the cassettes against the cores as shown in the figure below Remove the gel cassettes from the Buffer Cores and lay the gel cassettes well side up on a flat surface such as the benchtop Allow one edge to hang 1 cm over the side of the benchtop Carefully insert the Gel Knife s included with the Midi Cell beveled edge into the narrow gap between the two plates of the cassette Caution Use caution while inserting the Gel Knife between the two plates to avoid excessive pressure towards the gel Push up and down gently on the knife s handle to separate the plates You will hear a cracking sound indicating that you have broken the bonds which hold the plates together Repeat on each side of the cassette until the plates are completely separated Upon opening the cassette t
2. Amount of Buffer The amount of 1X Running Buffer required depends on the number of gels used for electrophoresis as described below Number of Gels Amount of Buffer 4 1 400 mL 3 1 250 mL 2 950 mL 1 750 mL Continued on next page General Guidelines for Preparing Samples and Buffers Continued Reducing Agent Important NuPAGE Antioxidant The NuPAGE Sample Reducing Agent contains 500 mM dithiothreitol DIT at a 10X concentration in a ready to use stabilized liquid form page 17 for ordering information and is used to reduce samples for electrophoresis B mercaptoethanol can be used with the Novex Midi gels at a final concentration of 2 5 v v Choice of the reducing agent is a matter of preference and either DTT or B mercaptoethanol can be used We recommend adding the reducing agent to the sample within an hour of loading the gel Avoid storing reduced samples for long periods of time even if they are frozen This will result in the reoxidation of samples during storage and produce inconsistent results Do not use the NuPAGE Antioxidant see below as a sample reducing agent The antioxidant is not efficient in reducing the disulfide bonds This will result in partially reduced bands with substantial background smearing in the lane The NuPAGE Antioxidant a proprietary reagent is added to the running buffer in the upper cathode buffer chamber only when performing electrophoresis of NuPAGE Novex Midi Ge
3. Flat Gel Loading Tips 200 pk LC1002 Gel Loading Tips Eppendorf Round 200 pk LC1010 Power Supply ZOOM Dual Power Supply 100 120 VAC 50 60 Hz 1 ZP10001 ZOOM Dual Power Supply 220 240 VAC 50 60 Hz 1 ZP10002 PowerEase 500 Power Supply 100 120 VAC 1 EI8600 50 60 Hz PowerEase 500 Power Supply 220 240 VAC 1 EI8700 50 60 Hz Novex Power Supply Adapters 1 set ZA10001 Technical Support Web Resources Visit the Invitrogen website at www invitrogen com for e Technical resources including manuals vector maps and sequences application notes SDSs FAQs formulations citations handbooks etc e Complete technical support contact information e Access to the Online Catalog e Additional product information and special offers Contact Us For more information or technical assistance call write fax or email Additional international offices are listed on our website www invitrogen com Corporate Headquarters Japanese Headquarters European Headquarters 5791 Van Allen Way LOOP X Bldg 6F Inchinnan Business Park Carlsbad CA 92008 USA 3 9 15 Kaigan 3 Fountain Drive Tel 1 760 603 7200 Minato ku Tokyo 108 0022 Paisley PA4 9RF UK Tel Toll Free 1 800 955 6288 Tel 81 3 5730 6509 Tel 44 0 141 814 6100 Fax 1 760 602 6500 Fax 81 3 5730 6519 Tech Fax 44 0 141 814 6117 E mail tech_support invitrogen com E mail jpinfo invitrogen com E mail eurotech invitrogen com SDS Safety Data Sheets S
4. The XCell4 SureLock Midi Cell is manufactured in the USA from domestic and International components TM To ensure safe reliable operation always operate the XCell4 SureLock Midi Cell according to instructions provided in this manual Wear protective gloves and safety glasses when working in a laboratory environment Invitrogen is not responsible for injuries or damages caused by improper use The symbols used on the XCell4 SureLock Midi Cell are explained below Used on the XCell4 SureLock Midi Cell to indicate an area where a potential shock hazard may exist Used on the XCell4 SureLock Midi Cell to indicate a warning The operator s manual should be consulted to avoid possible personal injury or instrument damage WEEE Waste Electrical and Electronic Equipment symbol indicates that this product should not be disposed of in unsorted municipal waste Follow local municipal waste ordinances for proper disposal provisions to reduce the environmental impact of WEEE Visit www invitrogen com weee for collection and recycling options Sicherheits Anweisungen Sicherheits Anweisungen SymbolerklArung A AN A TM Die XCell4 Elektrophorese Zelle muss mit einer geerdeten Gleichstrom Spannungsquelle betrieben werden die speziell f r die Elektrophorese entwickelt wurde Um eine Gef hrdung des Benutzers durch Stromschlag zu minimieren muss das Gleichstromversorgungsger t Gleichstromhochspannung produzieren die
5. output limit of the power supply Run fewer gels or use a power supply with higher power output Current reading on power supply is zero or very low Tape left on the bottom of the cassette Remove tape from the bottom of the cassette Connection to power supply not complete Check all connections with a voltmeter for conductance Insufficient buffer level Make sure the buffer in the Upper Buffer Chamber cathode is covering the wells Be sure there is sufficient buffer up to the fill line in the Lower Buffer Chamber Buffer is leaking from the Upper Buffer Chamber If the level of running buffer drops the electrophoresis core and cassettes are not properly seated Repeat the assembly see page 7 Check to ensure the XCell4 SureLock Assembly is in the locked position Run is faster than normal with poor resolution Buffers are too concentrated or incorrect Check buffer recipe dilute or re make if necessary Voltage current or wattage is set at a higher limit Decrease power conditions to the recommended run conditions page 13 16 Appendix Product Specifications XCell4 SureLock Midi Cell Specifications CE Gel Specifications Sample Well Capacity Maintenance Dimensions 7 5 1 x 8 35 w x 6 3 h inches Upper Buffer Chamber Volume 175 mL in each upper buffer chamber Lower Buffer Chamber Volume 700 mL with 4 gels Lower Buffer Chamb
6. DSs are available on our website at www invitrogen com sds Certificate of The Certificate of Analysis provides detailed quality control and product Analysis qualification information for each product Certificates of Analysis are available on our website Go to www invitrogen com support and search for the Certificate of Analysis by product lot number which is printed on the box 19 Purchaser Notification Limited Warranty 20 Invitrogen a part of Life Technologies Corporation is committed to providing our customers with high quality goods and services Our goal is to ensure that every customer is 100 satisfied with our products and our service If you should have any questions or concerns about an Invitrogen product or service contact our Technical Support Representatives All Invitrogen products are warranted to perform according to specifications stated on the certificate of analysis The Company will replace free of charge any product that does not meet those specifications This warranty limits the Company s liability to only the price of the product No warranty is granted for products beyond their listed expiration date No warranty is applicable unless all product components are stored in accordance with instructions The Company reserves the right to select the method s used to analyze a product unless the Company agrees to a specified method in writing prior to acceptance of the order Invitrogen makes every effort t
7. Invitrogen by technologies XCell4 SureLock Midi Cell For leak free electrophoresis of 1 4 midi gels Catalog no WR0100 Rev Date 22 June 2010 Manual Part no 25 0868 MANDO000534 ii Table of Contents RE Oe iii Safety Information sr oos ere EIER ee ed iv Sicherheits Anweisungen eee dee eere bane SEENEN trei ti ede rni eges vi Instructions de securite eene ee a e e e d d n uH i HR gun vii Experienced Users Procedure ss viii Contents ati osi ets nee ovili deo eid pe temo ebd sg a x Introduction EEE 1 Product Descriptioti sisi Mena eo ra ba ne D E EH ra a ir Pe e Has aere 1 Description EE 2 Experimental Overview este Ae e HU e ECRIRE EHE I e Lo HERE tienne lisent rires 4 Luigi pe Rs eenen 5 General Guidelines for Preparing Samples and Buffers een 5 Assembling the XCell4 SureLock Midr Eeln een ana 7 Performing Electrophoresis strates Gens aA baanden Raa e EERE Ea EEEa 13 Disassembling the XCell4 Surelock Mid Cell a ed 14 TroubleshoOtrigas uo ooo ede fee o npt epit dtd te oth fiiere irt ood 16 Appendix autas cie pan SERRE A RER DRM RA Sad o EORR UG Ve RUN uS Re ER 17 Product Specifications aue diit tete edite fne ieri i e ipee ae th ask MER 17 Accessory ne EE 18 Technical Supports 2 32 32 eme PUSH UR En mete eet ne e ees 19 Purchaser NotifiCati n ise eodem a ed apnd itte ep ia aa 20 iii Safety Information Safety TM During operation use the XCell4 SureLock Midi Cell with an external DC p
8. To obtain the best results and promote a uniform running of the stacking front load sample buffer in all the wells whether or not they contain samples B Pi ipet with sample Ei C KH a a Running c 4 Buffer Bet ad SOS CE CAT IT IT IT IT IT tr IQ Sample Sample m i iii ti it it tt Well Do not add any buffer or water to the Displacement Dam Important However if you have accidentally added any water or buffer to the Displacement Dam it does not affect the electrophoresis run Continued on next page 11 Assembling the XCell4 SureLock Midi Cell continued Adding Buffer into 1 Add the appropriate amount of 1X running buffer as described below to the Lower Buffer the Lower Buffer Chamber anode by pouring into the center of the Midi Chamber Cell over the XCell4 SureLock Assembly Fill to the fill level marked on the outside of the Midi Cell Number of Gels 1X Running Buffer 4 700 mL 3 680 mL 2 560 mL 1 540 mL Do not handle the lid if the cables are plugged into the power supply 2 Place the lid on the assembled XCell4 SureLock Midi Cell The lid will firmly seat if the and electrodes are properly aligned If the lid is not properly seated no power will go through the Midi Cell 3 With the power off connect the electrode cords to power supply red to jack black to jack Make sure the power is off before connecting the XCell4 SureLock M
9. ase The XCell4 SureLock Assembly slides down over the protrusion in the Midi Cell base 2 Placeone cassette on each side of the Buffer Core for each of the two cores For each cassette the shorter well side of the cassette faces IN towards the buffer core The slot on the back of the cassette must face out towards the lower buffer chamber figure below Buffer Core EI Notch faces in Second Gel or Buffer Dam Cassette Seno Slot faces out EE III z Side View 3 While holding the assembly together with your hands Figure A insert the Buffer Cores with the gel cassettes into the Lower Buffer Chamber such that the negative electrode fits into the opening in the gold plate on the Lower Buffer Chamber Figure B Always hold the cassette assembly by its edges as shown in the figure below Note If you are having difficulty inserting the assembly into the lower buffer chamber make sure the cathode black polarity indicator of the Buffer Core is aligned with the cathode black polarity indicator of the Lower Buffer Chamber Continued on next page Assembling the XCell4 SureLock Midi Cell continued Assembly with Procedure continued from previous page 4 Gels continued 4 The Upper Buffer Chamber cathode is the void formed between a gel and the Buffer Core at the center of each core 5 Lock the XCell4 SureLock Assembly by
10. e and gel over a container with the gel facing downward Gently push the gel knife through the slot in the cassette until the gel peels away from the plate Cut the foot off the gel after fixing and staining but before drying 10 Immediately stain the gel as desired Wash the XCell4 SureLock Midi Cell with a mild detergent and rinse with deionized water after each use If the wires on the Buffer Cores are damaged you can replace the wire using the spare wire available from Invitrogen page 17 For more information visit www invitrogen com or contact Technical Support page 19 Do not perform any repairs or service on the XCell4 SureLock Midi Cell to avoid damaging the Midi Cell 15 Troubleshooting Introduction Review the information below to troubleshoot your experiments using the XCell4 SureLock Midi Cell Problem Cause Solution Run taking longer than usual Buffers are too dilute Check buffer recipe re make if necessary Upper Buffer Chamber is leaking Make sure the buffer core is firmly seated If you are using the Buffer Dam make sure the dam is properly positioned in the core If the core gasket is damaged replace with a fresh gasket Check to ensure that the XCell4 SureLock Assembly is in the locked position Voltage is set too low Set correct voltage as listed on page 13 When running multiple gels the total current requirement exceeds the maximum
11. ension lever to the locked position indicated on the XCell4 SureLock Assembly This will squeeze the gels and Buffer Cores together creating leak free seals Continued on next page Experienced Users Procedure continued Add Buffer and Load Samples Disassembly Fill each of the Upper Buffer Chambers with 175 mL of the appropriate 1X running buffer For running NuPAGE Midi Gels under reducing conditions use 1X Running Buffer with 435 ul NuPAGE Antioxidant in each of the Upper Buffer Chambers Ensure that the Upper Buffer Chambers are not leaking Load the appropriate volume of the protein sample and molecular weight markers in the wells Add 700 mL 1X running buffer for 4 gels to the Lower Buffer Chamber anode by pouring into the center of the Midi Cell over the XCell4 SureLock Assembly Fill to the fill line marked on the Midi Cell Place the lid on the assembled XCell4 SureLock Midi Cell The lid firmly seats if the and electrodes are properly aligned With the power off connect the electrode cords to power supply Turn on the power and perform electrophoresis as follows Gel Type Voltage Run Time NuPAGE Bis Tris SDS PAGE with 200 V Constant 40 minutes MES Running Buffer NuPAGE Bis Tris SDS PAGE with 200 V Constant 55 minutes MOPS Running Buffer NuPAGE Tris Acetate SDS PAGE 150 V Constant 70 minutes denaturing reducing NuPAGE Tris Acetate SDS PAGE 150 V Constant 2 3 hours native Tr
12. er Material Polycarbonate Buffer Core Material Polycarbonate gold plated copper platinum plasticized silicone Buffer Dam Displacement Dam Material High density polyethylene Electrode Wire Platinum 0 010 diameter Electrical Limits 600 VDC or 200 Watts Operating Temperature 4 40 C TM The XCell4 SureLock Midi Cell is impervious to alcohol but not compatible with chlorinated hydrocarbons e g chloroform aromatic hydrocarbons e g toluene benzene or acetone The CE mark symbolizes that the product conforms to all applicable European Community provisions for which this marking is required Operation of the TM XCell4 SureLock Midi Cell is subject to the conditions described in this manual The protection provided by the equipment may be impaired if the equipment is used in a manner not specified by Invitrogen The XCell4 SureLock Midi Cell is compatible with NuPAGE Novex and Tris Glycine Midi precast gels The Novex Midi precast gels have the following specifications Cassette 15 cm x 10 3 cm Gel Size 13 cm x 8 3 cm Thickness 1 0 mm The maximum recommended sample volume well for different Novex Midi precast gels is listed below Gel 12242 well 20 well 26 well Thickness 1 0 mm 45 uL 25 uL 15 uL 15 uL marker Wash the XCell4 SureLock Midi Cell with a mild detergent and rinse with deionized water after each use 17 Accessory Products Additional Products 18 Additional
13. esteht Lesen Sie die Bedienungsanleitung um Verletzungen oder Besch digung des Ger tes zu vermeiden WEEE Instructions de securite Instructions de securite TM La XCell4 SureLock Midi Cell doit tre utiliser uniquement avec une alimentation en courant contimu DC externe et sp cialement concue pour en usage avec un systeme lectrophor se Cette alimentation doit tre isolee et sans contact avec la terre et doit etre equipee aven un systeme de protection en cas de Non Charge ou Circuit ouvert Le domaine d utilisation de la XCell4 SureLock Midi Cell est limit par les param tres de fonctionnement suivants Tension maximum 600 Volts DC Puissance maximum 200 Watts Temp rature 4 40 C TM La XCell4 SureLock Midi Cell est con ue de fa on ce que lorsque le couvercle de la cuve est ouvert la connection lectrique est interrompue Il ne faut en aucun cas essayer de faire fonctionner la cuve sans le couvercle La XCell4 SureLock Midi Cell satisfait aux normes de s curit EN 61010 1 La s curit est assur e dans tous les cas d une utilisation conforme au manuel d utilisation Dans le cas ou la cellule serait modifi e ou utilis e de mani re non sp cifi e dans le manuel d utilisation la s curit de l utilisateur pourrait tre remise en cause En cas d utilisation non conforme toutes les garanties sinsi que les certifications de conformit aux normes de s curit sont abroge s La responsabil
14. ffer Chambers assembled with gels with 175 mL of the appropriate 1X running buffer page 5 Use enough running buffer to completely cover the sample wells If using only the Displacement Dam there is no need to add any buffer to the Dam 2 Ifyouare running NuPAGE Novex Midi Gels under reducing conditions use 1X Running Buffer with 435 uL NuPAGE Antioxidant in each of the Upper Buffer Chambers Tip Slowly load the buffer into the Upper Buffer Chamber to avoid a lot of bubble formation Bubbles make sample loading more difficult Ensure that the Upper Buffer Chambers are not leaking If the level of running buffer drops the electrophoresis core and cassettes are not properly seated Disassemble the Buffer Core cassette assembly and repeat the assembly Continued on next page Assembling the XCell4 SureLock Midi Cell continued Loading Samples The recommended sample volume for each Novex Midi gel type is listed on page 17 1 Prepare your samples using the appropriate sample buffer at the desired protein concentration 2 Useapipette equipped with a sample loading tip page 17 to underlay the samples into the gel wells Lower the tip to the bottom of the sample well and slowly pipet sample into well without contaminating another well with the sample see figures A and B below The sample wells are marked on the cassette allowing you to easily identify the sample wells and load the samples properly into each well Note
15. h the XCell4 Lid produces an enclosed unit for electrophoresis The unique design of the XCell4 Buffer Core with the XCell4 SureLock Assembly provides a consistent seal without the use of any clamps or grease The Buffer Core is specifically designed to evenly dissipate heat resulting in consistent performance across all gels The Buffer Core contains the anode and cathode electrode assembly and has color coded electrical poles for easy intuitive assembly The XCell4 SureLock Midi Cell includes 2 Buffer Cores The XCell4 lid will fit onto the Midi Cell in only one orientation to prevent incorrect connection to the electrodes The Gel Knife is used to open the gel cassette to remove the gel b N Continued on next page Description of Parts Continued XCell4 Buffer Dam XCell4 Displacement Dam XCell SureLock Assembly Use the XCell4 Buffer Dam when you wish to run less than 4 gels in the XCell4 SureLock Midi Cell The Buffer Dam is placed in the I Buffer Core instead of a second gel This SIDE FACES Instructions for using the Buffer Dam to run CORE 1or 3 Midi Gels are provided on page 9 Use the XCell4 Displacement Dam when you wish to run less than 3 gels in the XCell4 SureLock Midi Cell The Displacement Dam is placed in the Lower Buffer Chamber instead of a second Buffer Core Instructions for using the Displacement Dam to run gt 1
16. he gel may adhere to either side Remove and discard the plate without the gel allowing the gel to remain on the other plate Note The Cassette Post small plastic piece near the top of the cassette may remain on either plate of the cassette after opening the two plates of the cassette The Cassette Post is designed to maintain proper electrophoresis conditions that result in optimal separation and does not interfere with the sample loading or electrophoresis run If blotting proceed to the western transfer protocol refer to the Novex Midi Gel System manual for more details Continued on next page Disassembling the XCell4 SureLock Midi Cell Continued Procedure continued Cleaning and Maintenance 9 If staining remove the gel from the cassette plate by one of the two methods e If the gel remains on the shorter notched plate use the sharp edge of the Gel Knife to remove the bottom foot of the gel if desired Hold the Gel Knife at a 90 angle to the gel and the slotted cassette plate Push straight down on the knife to cut the gel Repeat the motion across the gel to cut the entire foot Hold the cassette plate and gel over a container with the gel facing downward Use the knife to carefully loosen one lower corner of the gel and allow the gel to peel away from the plate 4 Gel knife 90 angle R Pre cast gel Gel foot Cassette plate e If the gel remains on the longer slotted plate hold the cassette plat
17. idi Cell to the power supply 4 Turn on the power See next page for electrophoresis conditions 12 Performing Electrophoresis Electrophoresis Conditions Once you have assembled the XCell4 SureLock Midi Cell and loaded your samples you are ready to perform electrophoresis You will need a power supply designed for electrophoresis page 4 The electrophoresis conditions for different Novex Midi Gels using the XCell4 SureLock Midi Cell are listed below Running Buffer native electrophoresis End 15 20 mA Gel Type Voltage Expected Current Run Time NuPAGE Bis Tris SDS PAGE denaturing 200 V Constant Start 160 200 mA 40 minutes reducing with MES Running Buffer End 120 170 mA NuPAGE Bis Tris SDS PAGE denaturing 200 V Constant Start 160 200 mA 55 minutes reducing with MOPS Running Buffer End 120 170 mA NuPAGE Tris Acetate SDS PAGE 150 V Constant Start 70 90 mA 70 minutes denaturing reducing End 50 60 mA NuPAGE Tris Acetate SDS PAGE native 150 V Constant Start 40 45 mA 2 3 hours End 15 20 mA Tris Glycine with Tris Glycine SDS Running 125 V Constant Start 40 50 mA 105 minutes Buffer End 20 25 mA Tris Glycine with Tris Glycine Native 125 V Constant Start 35 40 mA 1 12 hours Current readings are per gel At the end of the run turn off the power and disconnect cables from the power supply Proceed immediately to Disassembling the XCell4 SureLock Midi
18. ing the sample buffer and running buffer for each gel type is provided in the instruction manual included with each gel type This manual is available for downloading from our web site at www invitrogen com or by contacting Technical Support page 19 Recommended The recommended running buffer and sample buffer for each Novex Midi Buffers Gel is listed below You will prepare your sample in the appropriate sample buffer such that the final concentration of the sample buffer is 1X You need to dilute the running buffer to 1X final concentration before use See page 17 for ordering information on pre mixed buffers Gel Type Running Buffer Sample Buffer NuPAGE Novex NuPAGE MES SDS Running Buffer NuPAGE LDS Sample Buffer 4X Bis Tris Midi Gels 20X to resolve small proteins NuPAGE MOPS SDS Running Buffer NuPAGE LDS Sample Buffer 4X 20X to resolve mid size proteins NuPAGE Novex Tris Acetate Midi Gels NuPAGE Tris Acetate SDS Running NuPAGE LDS Sample Buffer 4X Buffer 20X to resolve large proteins Tris Glycine Native Running Buffer 10X Tris Glycine Native Sample Buffer 2X to perform native gel electrophoresis Novex Tris Glycine Midi Gels Tris Glycine SDS Running Buffer 10X to Tris Glycine SDS Sample Buffer 2X perform denaturing gel electrophoresis Tris Glycine Native Running Buffer 10X Tris Glycine Native Sample Buffer 2X to perform native gel electrophoresis
19. is Glycine with SDS Running Buffer 125 V Constant 105 minutes denaturing reducing Tris Glycine with Tris Glycine Native 125 V Constant 1 12 hours Running Buffer native electrophoresis Remove the lid Discard the buffer Unlock the XCell SureLock Assembly by moving the tension lever to the unlocked position indicated on the XCell SureLock Assembly Remove the Buffer Cores with gel cassettes from the Lower Buffer Chamber while holding the cassettes against the cores Remove the gel cassettes from Buffer Cores and carefully insert the Gel Knife s included with the Midi Cell beveled edge into the narrow gap between the two plates of the cassette Caution Use caution while inserting the Gel Knife between the two plates to avoid excessive pressure towards the gel Push up and down gently on the knife s handle to separate the plates Repeat on each side of the cassette until the plates are completely separated Upon opening the cassette remove and discard the plate without the gel allowing the gel to remain on the other plate If blotting proceed to the western transfer protocol refer to XCell II Blot Module manual for more details For staining remove the bottom foot of the gel with a Gel Knife if desired Hold the cassette plate and gel over a container with the gel facing downward Use the knife to carefully loosen one lower corner of the gel and allow the gel to peel away from the plate Contents C
20. it de Invitrogen ne p ut tre mise en cause en cas d accident et dommages caus s par une utilisation non conforme de la XCell4 SureLock Midi Cell Toute intervention de service ou de r paration doit tre unquement effectu e par Invitrogen ou par un agent agr e de Invitrogen La XCell4 SureLock Midi Cell est classifi par rapport la norme d isolation lectrique en classe II Signification des pictogrammes A AN A TM Sur le XCell4 SureLock Midi Cell signale les zones o un risque de choc lectrique peut exister TM Sur XCell4 SureLock Midi Cell signale un risque potentiel pour l utilisateur ou pour l equipement Veuillez consulter le mode d emploi WEEE vii Experienced Users Procedure Introduction A brief experienced users procedure for performing electrophoresis with the XCell4 SureLock Midi Cell is described below If you are a first time user of the XCell4 SureLock Midi Cell refer to the detailed protocols in this manual Prepare Sample and Buffer Prepare Gel Cassette Assembly with 4 Gels viii Prepare protein sample at the appropriate concentration using the NuPAGE LDS Sample Buffer or Tris Glycine SDS Sample Buffer for denaturing SDS PAGE and using the Tris Glycine Native Sample Buffer for native PAGE for use with NuPAGE Midi Tris Acetate Midi and Tris Glycine Midi Gels only Prepare the following 1X Running Buffer see page 5 for the amount of buffer needed based
21. ls under reducing conditions The NuPAGE Antioxidant migrates with the proteins during electrophoresis preventing the proteins from reoxidizing and maintaining the proteins in a reduced state The NuPAGE Antioxidant also protects sensitive amino acids such as methionine and tryptophan from oxidizing The NuPAGE Antioxidant is NOT compatible with gel systems other than the NuPAGE system as the antioxidant is not efficient at higher pH of other gel systems including Novex Tris Glycine Midi Gels If you are performing electrophoresis of NuPAGE Novex Midi Gels under reducing conditions using the XCell 4 SureLock Midi Cell use 175 mL 1X Running Buffer with 435 ul NuPAGE Antioxidant in each of the Upper Buffer Chambers Assembling the XCell4 SureLock Midi Cell Materials Needed e Appropriate precast gels page 17 e Running buffer 1X e Protein sample and protein standard e Gel loading tips Using the XCell4 The Buffer Cores and lid of the XCell4 SureLock Midi Cell are supplied SureLock Midi assembled Cell for the First If you are using the XCell4 SureLock Midi Cell for the first time you need to Time disassemble the XCell4 SureLock Midi Cell and wash various parts before use as described below 1 Remove the lid 2 Remove the Buffer Dam Displacement Dam and 2 Buffer Cores from the Lower Buffer Chamber 3 Wash various parts of the XCell4 SureLock Midi Cell with a mild detergent and rinse with dei
22. moving the tension lever to the locked position indicated on the XCell4 SureLock Assembly This will squeeze the gels and Buffer Cores together creating leak free seals 6 Proceed to loading samples and buffers pages 10 12 Assembly with Instructions are provided below to assemble the XCell4 SureLock Midi Cell with Less than 4 Gels less than 4 gels using the Buffer Dam and Displacement Dam included with the Midi Cell Note Do not use the XCell SureLock Buffer Dam with the XCell4 SureLock Midi Cell Gel Number Instructions Assembling Insert the XCell4 SureLock Assembly in its unlocked position with 3 gels into the Midi Cell The XCell4 SureLock Assembly should rest on the bottom of the Lower Buffer Chamber Assemble 1 Buffer Core with 2 gel cassettes and insert the Buffer Core gel cassette assembly into chamber 1 of the Lower Buffer Chamber as described on page 8 Assemble the second Buffer Core with a gel cassette and Buffer Dam see adjacent figure and insert into chamber 2 of the Lower Buffer Chamber Lock the XCell4 SureLock Assembly by moving the tension lever to the locked position indicated on the XCell4 SureLock Assembly to create a leak free seal Assembling Insert the XCell4 SureLock Assembly in its unlocked position with 2 gels into the Midi Cell between the two Buffer Cores The XCell4 SureLock Assembly should rest on the bottom of the Lower Buffer Chamber Assemble 1 Buffer Core with 2 gel cas
23. o ensure the accuracy of its publications but realizes that the occasional typographical or other error is inevitable Therefore the Company makes no warranty of any kind regarding the contents of any publications or documentation If you discover an error in any of our publications report it to our Technical Support Representatives Life Technologies Corporation shall have no responsibility or liability for any special incidental indirect or consequential loss or damage whatsoever The above limited warranty is sole and exclusive No other warranty is made whether expressed or implied including any warranty of merchantability or fitness for a particular purpose 2010 Life Technologies Corporation All rights reserved The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners 6 invitrogen by Lefe technologies Corporate Headquarters 5791 Van Allen Way Carlsbad CA 92008 T 1 760 603 7200 F 1 760 602 6500 E tech_support invitrogen com For country specific contact information visit our web site at www invitrogen com
24. on the number of gels used NuPAGE Bis Tris Midi Gels 1X NuPAGE MES SDS Running Buffer to resolve small proteins 1X NuPAGE MOPS SDS Running Buffer to resolve mid size proteins NuPAGE Tris Acetate Midi Gels 1X NuPAGE Tris Acetate SDS Running Buffer to resolve large proteins 1X Tris Glycine Native Running Buffer to perform native gel electrophoresis Tris Glycine Midi Gels 1X Tris Glycine SDS Running Buffer for denaturing SDS PAGE 1X Tris Glycine Native Running Buffer for native gel electrophoresis Remove the gel cassette from the pouch and rinse with deionized water Peel off the tape covering the slot on the back of the gel cassette and pull the comb out of the cassette Rinse the wells with 1X running buffer and fill the sample wells with running buffer Instructions for assembling four gels are described below For assembling less than four gels see page 9 1 Insert the XCell4 SureLock Assembly in its unlocked position into the center of the Midi Cell base The XCell4 SureLock Assembly slides down over the protrusion in the Midi Cell base Place one cassette on each side of both Buffer Cores While holding the assembly together with your hands insert the Buffer Cores with the gel cassettes into the Lower Buffer Chamber such that the negative electrode fits into the opening in the gold plate on the Lower Buffer Chamber Always hold the cassette assembly by its edges TM Lock the XCell4 SureLock Assembly by moving the t
25. onized water before use Do not damage the wires on the core during the washing To avoid contamination from possible residual acrylamide wear gloves and laboratory coat when loading and unloading gels into the XCell4 SureLock Midi Cell and when staining destaining or drying gels Preparing Gel Cassettes Cut open the gel cassette pouch and drain away the gel packaging buffer Remove the gel cassette from the pouch and rinse with deionized water Peel off the tape covering the slot on the back of the gel cassette In one fluid motion pull the comb out of the cassette Quom o mop Use a pipette to gently wash the cassette wells with 1X running buffer Invert the gel and shake to remove buffer Repeat twice Fill the sample wells with running buffer Be sure to displace all air bubbles from the cassette wells as they will affect sample running Note Always handle the cassette by its edges only To obtain the best results use gels of the same type and same percentage of Note acrylamide in the XCell4 SureLock Midi Cell Avoid mixing different gel types and percentage of gels Continued on next page Assembling the XCell4 SureLock Midi Cell continued Assembly with Instructions are provided below to assemble the XCell4 SureLock Midi Cell 4 Gels with 4 precast midi gels See page 9 to assemble with less than 4 gels 1 Insert the XCell4 SureLock Assembly in its unlocked position into the center of the Midi Cell b
26. ontents The components included with the XCell4 SureLock Midi Cell are listed below For more details on each component see page 2 Components Quantity XCell4 Lower Buffer Chamber 1 XCell4 Lid with Cables 1 XCell4 Buffer Cores 2 XCell4 Buffer Dam 1 XCell4 Displacement Dam 1 XCell4 SureLock Assembly 1 Gel Knife 1 Intended Use For research use only Not intended for any animal or human therapeutic or diagnostic use Introduction Product Description XCell4 SureLock Midi Cell System Overview Features The XCell4 SureLock Midi Cell s unique design allows you to quickly and easily perform simultaneous vertical electrophoresis of 1 4 midi gels without leaking resulting in consistent performance The XCell4 SureLock Midi Cell is built with tough plastic material to boost durability and features a specially designed lid to enhance user safety XCell4 SureLock Midi Cell Buffer Core Banana Plug Lower Buffer Chamber The XCell4 SureLock Midi Cell consists of a Lower Buffer Chamber two Buffer Cores Buffer Dam Displacement Dam and a lid with cables see next page for detailed description of parts Electrophoresis of proteins on midi gels in the XCell4 SureLock Midi Cell is performed after assembling each Buffer Core with 2 midi gels or a Buffer Core with a midi gel and a Buffer Dam The Buffer Core gel assembly is inserted into the Lower Buffer Chamber and is sealed using a positive locking mechani
27. or 2 midi gels are provided on page 9 Do not add any buffer or water to the Displacement Dam TM The positive locking action of the XCell SureLock Assembly ensures trouble free leak free electrophoresis of protein gels Move the lever on the XCell SureLock Assembly to a locked position to generate an even horizontal force which seals the gel cassette Buffer Core assembly firmly into position in the Lower Buffer Chamber The locked and unlocked positions are marked shown with an arrow in figure A below on the XCell SureLock Assembly for easy intuitive assembly Figure B shows the markings in detail Note Do not use the Gel Tension Wedge supplied with the XCell SureLock Mini Cell with the Midi Cell as the wedge is specifically designed for the Mini Cell and will not fit correctly in the Midi Cell Figure A Figure B The locked and unlocked positions marked on the assembly are shown in detail below lock unlock Experimental Overview Experimental The table below outlines the experimental steps necessary to perform Outline electrophoresis using the XCell4 SureLock Midi Cell For more details on each step see indicated pages Prepare protein samples Prepare running buffer Assemble the XCell4 SureLock Midi Cell with the desired number of gel cassettes Load the 1X Running Buffer into the upper buffer chamber Load protein samples into the wells of the gel Load running buffer into the Lo
28. ower supply designed specifically for electrophoresis applications This power supply must be isolated from ground so that the DC output is floating and must be equipped with No Load or Open Circuit protection The maximum electrical TM operating parameters for the XCell4 SureLock Midi Cell are Maximum Voltage Limit 600 Volts DC Maximum Power Limit 200 Watts Operating Temperature 4 40 C The XCell4 SureLock Midi Cell lid is designed such that if the lid is removed the electrical connection to the Midi Cell will be broken Do not attempt to use the cell without the cell lid The XCell4 SureLock Midi Cell is designed to meet EN61010 1 Safety Standards This product is safe to use when operated in accordance with this instruction manual If this Midi Cell is used or modified in a manner not specified in this manual then protection afforded by the Midi Cell will be impaired Alteration of this Midi Cell will e Void the warranty e Void the EN61010 1 safety standard certification e Create a potential safety hazard Invitrogen is not responsible for any injury or damage caused by use of this Midi Cell when operated for purposes which it is not intended All repairs and service should be performed by Invitrogen TM The XCell4 SureLock Midi Cell is classified as Class II for protection against electrical shock Continued on next page Safety Information continued Important Informational Symbols A A A
29. reagents that may be used for electrophoresis of proteins are available separately from Invitrogen Ordering information is provided below For details visit www invitrogen com or contact Technical Support page 19 A large variety of Novex Midi Gels and protein standards are available for use with the Midi Cell see www invitrogen com for details Midi Cell Spare Parts Quantity Catalog no XCell4 SureLock Midi Cell Lid 1 WRO101 XCell4 SureLock Midi Cell Lower Buffer Chamber 1 WR0102 XCell4 SureLock Midi Cell SureLock Assembly 1 WR0103 XCell4 SureLock Midi Cell Buffer Core 1 WRO104 XCell4 SureLock Midi Cell Buffer Dam 1 WR0105 XCell4 SureLock Midi Cell Displacement Dam 1 WR0106 XCell4 SureLock Midi Cell Wire 1 WR0107 XCell4 SureLock Midi Cell Gasket 1 WR0108 Pre Mixed Buffers NuPAGE LDS Sample Buffer 4X 10 mL NP0007 NuPAGE Sample Reducing Agent 10X 250 uL NP0004 10 mL NP0009 NuPAGE Antioxidant 15 mL NP0005 NuPAGE MOPS SDS Running Buffer 20X 500 mL NP0001 NuPAGE MES SDS Running Buffer 20X 500 mL NP0002 NuPAGE Tris Acetate SDS Running Buffer 20X 500 mL LA0041 Novex Tris Glycine Native Running Buffer 10X 500 mL LC2672 Novex Tris Glycine Native Sample Buffer 2X 20 mL LC2673 Novex Tris Glycine SDS Running Buffer 10X 500 mL LC2675 Novex Tris Glycine SDS Sample Buffer 2X 20mL LC2676 Pipette Tips Gel Loading Tips Standard Round 200 pk LC1001
30. settes and insert the Buffer Core gel cassette assembly into chamber 1 of the Lower Buffer Chamber as described on page 8 Insert the Displacement Dam into chamber 2 of the Lower Buffer Chamber figure A Lock the XCell4 SureLock Assembly by moving the tension lever to the locked position indicated on the XCell4 SureLock Assembly to create a leak free seal figure B Continued on next page Assembling the XCell4 SureLock Midi Cell continued Assembly with Less than 4 Gels continued TM Insert the XCell4 SureLock Assembly in its unlocked position into the Midi Cell between the two Buffer Cores The XCell4 SureLock Assembly should rest on the bottom of the Lower Buffer Chamber Assemble 1 Buffer Core with 1 gel cassette and 1 Buffer Dam and insert the Assembling with 1 gel Adding Buffer into the Upper Buffer Chamber 10 assembly into chamber 1 of the Lower Buffer Chamber as described on page 8 Insert the Displacement Dam into chamber 2 of the Lower Buffer Chamber as described on the previous page Lock the XCell4 SureLock Assembly by moving the tension lever to the locked position indicated on the XCell4 SureLock Assembly to create a leak free seal After assembling the XCell4 SureLock Midi Cell load buffer in the Upper Buffer Chamber as described below The Upper Buffer Chamber cathode is the void formed between a gel and Buffer Core at the center of each core 1 Filleach of the Upper Bu
31. sm that seals the Buffer Core gel assembly firmly into position and ensures leak free electrophoresis The Buffer Core gel assembly creates separate upper buffer chambers for each gel and a common lower buffer chamber shared by all gels The lid is placed onto the Midi Cell and fits on the Midi Cell in only one orientation to prevent incorrect connection to the electrodes TM The unique design of the XCell4 SureLock Midi Cell evenly dissipates heat resulting in consistent performance of all gels without the need for any external cooling device The Buffer Core electrical poles are color coded for easy intuitive assembly Important features of the XCell4 SureLock Midi Cell are listed below e User friendly design without any clamps or grease for leak free electrophoresis e Flexibility to perform electrophoresis of 1 4 midi gels e Unique heat dissipating design of the XCell4 SureLock Midi Cell produces consistent results without the need for any cooling device e Built in safety features to enhance user safety Description of Parts XCell4 Lower Buffer Chamber XCell4 Buffer Core XCell4 Lid with Cables Gel Knife The XCell4 Lower Buffer Chamber when assembled with the XCell4 SureLock Assembly and Buffer Cores creates two individual chambers for electrophoresis The Lower Buffer Chamber volume is 700 mL and is used as a heat sink and ion reservoir during electrophoresis The Lower Buffer Chamber when combined wit
32. vom Erdleiter elektrisch getrennt ist und es muss mit einer Schutzschaltung gegen Fehlstr me FI Schutzschalter ausgestattet sein Desweiteren sollten folgende TM Laufbedingungen f r die XCell4 Elektrophorese Zelle eingehalten werden Maximale Spannung 600 Volts DC Maximale Leistung 200 Watts Temperatur 4 40 C TM Beim Entfernen des Kammerdeckels der XCell4 Electrophorese Zelle wird der Stromflub unterbrochen Die Benutzung der Zelle ohne Deckel ist verboten Die XCell4 Elektrophorese Zelle entspricht den Sicherheits Richtlinien EN 61010 1 Es besteht keinerlei Gef hrdung f r den Benutzer wenn die Richtlinien der Gebrauchsanweisung eingehalten werden Wird die Elektophorese Zelle in einer Art und Weise benutzt bzw modifiziert die nicht in der Gebrauchsanleitung beschrieben ist werden die Sicherheitvorkehrungen zum Schutz des Betreibers beeintr chtigt Bei unsachgem sser Handhabung entfallen alle Garantieanspr che und Sicherheitszulassungen EN61010 1 Invitrogen haftet nicht f r Verletzungen oder Sch den jeglicher Art wenn die Elektrophorese Zelle f r Anwendungen eingesetzt wird f r die die Zelle nicht entwickelt wurde Alle Reparaturen und Wartungen m ssen von Invitrogen oder einem von Invitrogen authorisierten H ndler vorgenommen werden Das Geh use der XCell4 Elektrophorese Zelle besteht aus Kunststoff und enth lt keine leitenden Teile Weist auf eine Zone hin wo Gef hrdung durch Stromschlag b
33. wer Buffer Chamber Perform electrophoresis using the recommended run conditions Disassemble the XCell4 SureLock Midi Cell Materials Needed Ordering information is on page 19 e Power supply see below e Novex Midi Gels e Gel loading tips e Protein samples e Protein Standard e Sample buffer next page e Running buffer next page TM Power Supply The XCell4 SureLock Midi Cell is used with an external DC power supply Requirements designed for electrophoresis This power supply must e Be isolated from the ground so that the DC output is floating and must be equipped with No Load or Open Circuit protection e Be able to operate at 200 V The ZOOM Dual Power Supply and PowerEase 500 Power Supply are compatible for use with Novex Midi Gels See page 19 for ordering information TM Note The XCell4 SureLock Midi Cell has covered power leads to be in compliance with the latest IEC International Electrotechnical Commission and EN European standards and reduce the risk of electrical shock due to exposure to bare metal To allow connection of the XCell4 SureLock Midi Cell covered power leads with certain power supplies use Novex Power Supply Adapters available from Invitrogen page 19 Methods General Guidelines for Preparing Samples and Buffers Introduction General guidelines for preparing samples and buffers for Novex Midi precast gels are discussed below Detailed instructions for prepar

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