XCell SureLock
Contents
1. Chamber to cover the slot at the bottom of the gel 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 ata higher limit Decrease power conditions to recommended running conditions see page 13 Cannot see the sample wells to load sample There is little contrast between the sample well and the rest of the gel Mark cassette at the bottom of the wells with a marker pen prior to assembling the Upper Buffer Chamber Illuminate the bench area with a light source placed directly behind the XCell TM SureLock unit 12 Appendix Buffers and Running Conditions Introduction The buffers and running conditions for the different Novex mini gels using the XCell SureLock Mini Cell are listed below For ordering information on buffers see page 15 Gel Type Sample Running Buffer Power Run Buffer Settings Time Tris Glycine Tris Glycine Tris Glycine SDS Voltage 90 minutes SDS PAGE SDS Sample Running Buffer 125 V Constant denaturing Buffer LC2676 LC2675 Expected Current DO Start 30 40 mA reducing End 8 12 mA Tris Glycine Add NuPAGE SDS PAGE Sample denaturing Reducing Agent reducing NP0004 Tris Glycine Tris Glycine Tris Glycine Native Voltage 1 12 hours Native Native Sample Running Buffer 125V Constant B2. XCell SureLock Mini Cell 1kit EI0001 SureLock Retrofit Kit for XCell II Mini Cell 1kit EI0020 XCell SureLock Mini Cell w XCell II Mini Cell 1 kit EI0002 XCell II Blot Module 1 EI9051 PowerEase 500 Power Supply 1 EI8600 Replacement Parts Quantity Cat no Lower Buffer Chamber 1 EI0013 Gel Tension Wedge 1 EI0011 Gel Knife 1 EI9010 Buffer Core with Electrodes 1 EI9014 Lid with Cables 1 EI0010 Buffer Dam 1 EI0012 Buffer Core Replacement Wire 2 EI9022 Gaskets for EIO001 and EI9051 2 EI9016 Repair Kit for Buffer Core includes wire wire 1 Kit EI9021 insulation post nut washer Pipette Tips for Gel Loading Quantity Cat no Gel Loading Tips Standard Round 200 pk LC1001 Flat Gel Loading Tips 200 pk LC1002 Gel Loading Tips Eppendorf Round 200 pk LC1010 Continued on next page 15 Related Products Continued Pre Mixed Buffers Quantity Cat no NuPAGE LDS Sample Buffer 4X 10 mL NP0007 NuPAGE Sample Reducing Agent 10X 250 uL NP0004 10mL 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 NuPAGE Transfer Buffer 20X 125 mL NP0006 Novex Tris Glycine SDS Running Buffer 10X 500 mL LC2675 Novex Tris Glycine SDS Sample Buffer 2X 20mL LC2676 Novex Tris Glycine Na
3. the appropriate running buffer see page 13 Use enough running buffer to completely cover the sample wells 5 Ensure that the Upper Buffer Chamber is not leaking If the level of running buffer drops the electrophoresis core and cassettes are not properly seated Repeat steps 6 and 7 on page 6 6 Use the pipette equipped with a round sample loading tip Cat no LC1001 to underlay the samples into the gel wells see figure below Lower the tip to the bottom of the sample well and slowly pipet sample into well without contaminating another well with the sample If you have difficulty distinguishing the sample wells when loading the sample see Troubleshooting Note 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 Pipetwih sample Y A N Z2 g f M Running c f CP Buffer espresse AAA ARRAS ft tt at ott ty it Pd Sample Sample min 11 LD Lt II el Continued on next page Gel Electrophoresis Protocol Continued Procedure 1 Fill the Lower Buffer Chamber anode by pouring 600 mL of running buffer through the gap between the Gel Tension Wedge and the back of the Lower Buffer Chamber as shown in the figure below Note If you are using Tricine and NuPAGE Gels we recommend that you fill the lower buffer chamber completely 600 mL as this will help dissipate heat during the run Fo
4. Knife 1 each Gel Tension Wedge 1 each Buffer Dam 1 each SureLock Mini Cell Lid 1 each EI0001 XCell IT Blot Module SureLock Retrofit Kit for XCell II Mini Cell XCell SureLock Mini Cell with XCell II Blot Module Kit Safety See page 19 for safety information and explanation of Information symbols used on the instrument Intended Use For research use only Not intended for any animal or human therapeutic or diagnostic use ii Product Specifications Product Description Mini Cell Lid Buffer Dam TM The XCell SureLock Mini Cell s unique design allows you to run mini gels quickly easily and leak free without any clamps or grease The set up time is only 30 seconds just drop the Buffer Core and the Gel Tension Wedge into the Lower Buffer Chamber insert the gels and pull the Gel Tension Wedge forward The perfect no leak seal results in no mess and consistent performance Tough polycarbonate construction of the unit boosts durability Retractable plugs recessed jacks and a specially designed lid enhance user safety You can transfer with the optional XCell II Blot Module Cat No EI9051 using the same Lower Buffer Chamber Buffer Core Lower Buffer Gel Chamber Tension Wedge Continued on next page Product Specifications Continued XCell Dimensions 11 x 12 x 16 cm SureLock Upper Buffer Chamber Capacity 200mL Mini Cel
5. art 30 40 mA End 8 12 mA IEF pH 3 7 IEF pH 3 7 IEF pH 3 7 Voltage 2 5 hours Sample Buffer Cathode Buffer 100V 1 hour LC5371 LC5370 200V 1 hour IEF Anode Buffer 500V 30 minutes LC5300 Expected Current Start 5 mA End 6 mA IEF pH 3 10 IEF pH 3 10 IEF pH 3 10 Voltage 2 5 hours Sample Buffer Cathode Buffer 100V 1 hour LC5811 LC5310 200V 1 hour IEF Anode Buffer 500V 30 minutes LC5300 Expected Current Start 5 mA End 6 mA TBE Hi Density TBE TBE Running Voltage 30 90 Sample Buffer Buffer LC6675 200V Constant minutes LC6678 Expected Current Start 10 18 mA End 4 6 mA TBE Urea TBE Urea TBE Running Voltage 50 75 Sample Buffer Buffer LC6675 180V Constant minutes LC6876 or Expected Current Prep TBE Urea Start 10 20 mA Sample Buffer d LC6877 End 6 14 mA DNA Hi Density TBE TBE Running Voltage 90 minutes Retardation Sample Buffer Buffer LC6675 at 100V Constant LC6678 1 2X concentration Expected Current Start 12 15 mA End 6 15 mA Current readings are per gel 14 Related Products Additional Many of the components of the XCell SureLock Mini Products Cell as well as additional 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 call Technical Support page 17 Apparatus Quantity Cat no
6. cturing 2 use of the product or its components to provide a service information or data 3 use of the product or its components for therapeutic diagnostic or prophylactic purposes or 4 resale of the product or its components whether or not such product or its components are resold for use in research For products that are subject to multiple limited use label licenses the terms of the most restrictive limited use label license shall control Life Technologies Corporation will not assert a claim against the buyer of infringement of patents owned or controlled by Life Technologies Corporation which cover this product based upon the manufacture use or sale of a therapeutic clinical diagnostic vaccine or prophylactic product developed in research by the buyer in which this product or its components was employed provided that neither this product nor any of its components was used in the manufacture of such product If the purchaser is not willing to accept the limitations of this limited use statement Life Technologies is willing to accept return of the product with a full refund For information about purchasing a license to use this product or the technology embedded in it for any use other than for research use please contact Out Licensing Life Technologies 5791 Van Allen Way Carlsbad California 92008 or outlicensing lifetech com Appendix B Safety Safety Information Safety During operation the XCell SureLock Mini Ce
7. e Sicherheits Anweisungen Sicherheits Anweisungen Die SureLock Elektrophorese Zelle mub mit einer geerdeten Gleichstrom Spannungsquelle betrieben werden die speziell f r die Elektrophorese entwickelt wurde Um eine Gef hrdung des Benutzers auszuschlieben sollten folgende Laufbedingungen eingehalten werden Maximale Spannung 1500 Volt Maximale Leistung 75 Watt Maximale Temperatur 70 C Beim Entfernen des Kammerdeckels der SureLock Electrophorese Zelle wird der Stromflub unterbrochen Die Benutzung der Zelle ohne Deckel ist verboten Die SureLock 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 Sicherheitvorkehrugen zum Schutz des Betreibers beeintr chtigt Bei unsachgem ber 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 SureLock Elektrophorese Zelle besteht aus Kunststoff und enth lt
8. el 1 760 603 7200 Tel Toll Free 1 800 955 6288 Fax 1 760 602 6500 E mail tech supportGinvitrogen com European Headquarters Inchinnan Business Park 3 Fountain Drive Paisley PA4 9RF UK Tel 44 0 141 814 6100 Tech Fax 44 0 141 814 6117 E mail eurotech invitrogen com 17 Purchaser Notification Limited Use Label License No 5 Invitrogen Technology 18 The purchase of this product conveys to the buyer the non transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer whether the buyer is an academic or for profit entity The buyer cannot sell or otherwise transfer a this product b its components or c materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes The buyer may transfer information or materials made through the use of this product to a scientific collaborator provided that such transfer is not for any Commercial Purpose and that such collaborator agrees in writing a not to transfer such materials to any third party and b to use such transferred materials and or information solely for research and not for Commercial Purposes Commercial Purposes means any activity by a party for consideration and may include but is not limited to 1 use of the product or its components in manufa
9. f the benchtop Carefully insert the Gel Knife s beveled edge into the narrow gap between the two plates of the cassette Note Do not push the knife forcefully between the cassette plates or you may cut into the gel Push up and down gently on the knife s handle to separate the plates You will hear a cracking sound which means you have broken the bonds which hold the plates together Repeat until you have broken the bonds on one side Rotate the cassette and repeat Steps 5 6 until the two plates are completely separated Upon opening the cassette the gel may adhere to either side 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 without removing the gel from the plate refer to IM 9051 for more details Note Trying to remove the gel at this point may result in the gel tearing Proceed to Step 11 Continued on next page Disassembling the XCell SureLock Mini Cell Continued Procedure 10 If staining remove the gel from the cassette plate continued 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 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 co
10. he gel loading wells 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 Note Be sure to displace all air bubbles from the cassette wells as they will affect sample running Lower the Buffer Core into the Lower Buffer Chamber so that the negative electrode fits into the opening in the gold plate on the Lower Buffer Chamber as shown in the figure L Buffer Core Lower Buffer Chamber Lower Buffer Chamber and Buffer Core Front View Continued on next page 5 Assembling the XCell SureLock Mini Cell Continued Procedure 5 Insert the Gel Tension Wedge into the XCell continued SureLock behind the buffer core Make sure the Gel Tension Wedge is in its unlocked position see next page allowing the wedge to slip easily into the XCell SureLock unit The Gel Tension Wedge should rest on the bottom of the Lower Buffer Chamber 6 Insert gel cassettes into the lower buffer chamber Place one cassette behind the core and one cassette in front of the core 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 see figure below Buffer Core a SI H Notch faces in nl Second Gel or Buffer Dam Ca
11. ilis 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 responsabilit de Invitrogen ne p ut tre mise en cause en cas d accident et dommages caus s par une utilisation non conforme de la Cellue d Electrophor se Toute intervention de service ou de r paration doit tre unquement effectu e par Invitrogen ou par un agent agr e de Invitrogen La Cellule d Electrophor se SureLock est classifi par rapport la norme d isolation lectrique IEC 536 en classe II Signification des pictogrammes Lx gt gt Sur le Cellule d Electrophor se XCell SureLock signale les zones o un risque de choc lectrique peut exister Sur le Cellule d Electrophor se XCell SureLock signale un risque potentiel pour l utilisateur ou pour l equipement Veuillez consulter le mode d emploi WEEE Waste Electrical and Electronic Equipment 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 A 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 v
12. ins platinum electrodes gold terminals and silicone gaskets Cat no EIO010 Continued on next page 3 Description of Parts Continued Gel Tension Cat no EI0011 Wedge The positive locking action of the Gel Tension Wedge ensures trouble free leak free electrophoresis of protein gels Push the lever on the Gel Tension Wedge forward into 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 Buffer Dam Cat no EI0012 Use the Buffer Dam when you need to run only one Novex gel Gel Knife Cat no EI9010 Assembling the XCell SureLock Mini Cell Removing Gel Cassette Procedure 1 Cut open the gel cassette pouch with scissors and remove cassette Drain away the gel packaging buffer Remove the gel cassette from the pouch and rinse with deionized water Note Always handle the cassette by its edges only Always wear protective clothing when performing laboratory experiments To avoid contamination from possible residual acrylamide wear protective gloves when loading and unloading the XCell SureLock Mini Cell and when preparing staining destaining and drying gels 1 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 This exposes t
13. invitrogen by technologies XCell SureLock Mini Cell For leak free electrophoresis of mini gels Catalog nos EIO001 EIO020 EIO002 Rev Date 6 May 2010 Manual part no IM 9003 MANDO000739 Table of Contents Kit Contents uen dte Fendt m ettet re datas ii Product Specificatiofis 2 22 ue dan Reese 1 Description of Parts ss 3 Assembling the XCell SureLock Mini Cell sse 5 Gel Electrophoresis Protocol ss 8 Disassembling the XCell SureLock Mini Cell 10 Troubl shoeotfig 5n eere rete peer epe te edes 12 Buffers and Running Conditions sse 13 Related Products etae ires een at reis 15 Technical Support aiii eei andi ue eile ete tia tee 17 Purchaser Notification 18 Safety Information 19 Sicherheits Anweisungen ss 21 Instt ctions ide securite senti ananas 22 Kit Contents Types of Kits This manual is supplied with the following kits Kit Cat no XCell SureLock Mini Cell EI0001 SureLock Retrofit Kit for XCell II Mini Cell EI0020 XCell SureLock Mini Cell with XCell II Blot Module EI0002 Kit Kit The components included with the different kits Components arelisted below See page 3 for more details Kit Components XCell SureLock Mini Cell Gel Tension Wedge 1 each Buffer Dam 1 each Lower Buffer Chamber 1 each Buffer Core 1 each Mini Cell Lid 1 each Gel
14. isit our web site at www invitrogen com
15. k is rated at 1500 VDC 75 Watts but for running protein gels the maximum voltage wattage recommended is 500 VDC 50 Watts Continued on next page 19 Safety Information Continued Symbols on Instrument Informational Symbols A A 2 Important 20 The following table describes the symbols displayed on the instrument The symbols used on the XCell SureLock Mini Cell are explained below Used on the XCell SureLock Mini Cell to indicate an area where a potential shock hazard may exist Used on the XCell SureLock Mini 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 The XCell SureLock Mini Cell is manufactured in the USA from domestic and International components To ensure safe reliable operation always operate the XCell SureLock Mini Cell according to instructions provided in this manual Wear protective gloves and safety glasses when working in a laboratory environment Invitrogen products are intended for in vitro use only Invitrogen is not responsible for injuries or damages caused by improper us
16. keine leitenden Teile Die QuickPoint Elektrophorese Zelle ist gegen Stromschlag nach der IEC 536 Norm in Klasse II eingestuft Symbolerkl rung Lx gt e Weist auf eine Zone hin wo Gef hrdung durch Stromschlag besteht Lesen Sie die Bedienungsanleitung um Verletzungen oder Besch digung des Ger tes zu vermeiden WEEE Waste Electrical and Electronic Equipment 21 Instructions de securite Instructions de securite La Cellule d Electrophor se SureLock doit tre uniquement une utilisation avec une alimentation en courant contimu DC externe et sp cialement con ue pour en lectrophor se Cette alimentation doit tre avec s paration galvanique de facon que la sortie DC du secondaire ai un potentiel lectrique flottant Le domaine d utilisation de la Celleule d Electrophor se SureLockTM est limit par les param tres de fontctionnement suivants Tension maximum 1 500 V continu Puissance maximum 75W Temp rature maximum 70 C La Cellule d Electrophor se SureLock est con ue de fa on 4 ce que lorsque le couvercle de la cuve est ouvert la connection lectrique avec l unit est interrompue Il ne faut en aucun cas essayer de faire fonctionner la cuve sans le couvercle La Cellule d Electrophor se SureLock 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 ut
17. l Nn 2 Lower Buffer Chamber Capacity 600 mL Specifications Material Polycarbonate Electrode Wire Platinum 0 010 diameter Electrical Limits 1 500 VDC or 75 Watts Temperature Limit 70 C The XCell SureLock Mini Cell is impervious to alcohol but not compatible with chlorinated hydrocarbons e g chloroform aromatic hydrocarbons e g toluene benzene or acetone Gel Gel Cassette 10 cm x 10cm Specifications Thickness 1 0 mm or 1 5 mm Gel Size 8 cm x 8 cm Combs 1 well 2D well 5 well 9 well 10 well 15 well and 17 well Sample Well The maximum recommended sample volume well Capacity for different Novex mini gels is listed below For additional details on loading volumes and maximum protein load refer to the Novex Pre Cast Gel Electrophoresis Guide or the NuPAGE Technical Guide Gel 1 well 2D 5 well 9 well 10 well 15 well 17 well Thickness well 15 uL 1 0 mm 700 pL 400 uL 60 uL 28 uL 25 uL 15 uL 1 5 mm N A 600 uL N A N A 37uL 25 uL 25 uL Description of Parts Introduction Lower Buffer Chamber Buffer Core Cell Safety Lid with Cables The various parts included with the XCell SureLock TM Mini Cell are described below See page 15 for ordering information Cat no EIO013 The Lower Buffer Chamber has a capacity of approximately 800 mL of buffer and is used as a heat sink during the electrophoresis of proteins Cat no EI9014 Buffer Core conta
18. ll must be used with an external DC power supply designed specifically for electrophoresis applications This power supply must be isolated from ground so that the DC output is floating The PowerEase 500 Programmable Power Supply page 15 meets these requirements The maximum electrical operating TM parameters for the XCell SureLock Mini Cell are Maximum Voltage Limit 1500 VDC Maximum Power Limit 75 Watts Maximum Operating Temperature Limit 70 C The XCell SureLock Mini Cell s lid is designed such that if the lid is removed the electrical connection to the unit will be broken Do not attempt to use the cell without the cell lid Do not use lids from other mini cells The XCell SureLock Mini 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 unit is used or modified in a manner not specified in this manual then protection afforded by the unit will be impaired Alteration of this unit 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 unit when operated for purposes which it is not intended All repairs and service should be performed by Invitrogen The XCell SureLock Mini Cell is classified as Class II of IEC 536 for protection against electrical shock The XCell SureLoc
19. ntainer 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 a Gel knife 90 angle a Precast gel Gel foot Cassette plate e If the gel remains on the longer slotted plate hold the cassette plate 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 11 Immediately fix stain or transfer the gel as desired Maintenance Wash the XCell SureLock Mini Cell with a mild detergent and rinse with deionized water after each use 11 Troubleshooting Problem Cause Solution Run taking Buffers are too dilute Check buffer recipe remake longer than if necessary usual Upper buffer chamber is leaking Make sure the buffer core is firmly seated the gaskets are in place and the gel tension lever is locked Voltage is set too low Set correct voltage Current reading on power supply is zero or very low Tape left on the bottom of the cassette Remove tape from bottom of cassette Connection to power Check all connections with a supply not complete voltmeter for conductance Insufficient buffer Make sure the upper buffer level cathode is covering the wells of the gel Be sure there is sufficient buffer in the Lower Buffer
20. r other gel types a filled lower buffer chamber is recommended but not required The XCell SureLock Mini Cell will function as long as the lower buffer chamber is filled enough to cover the slot at the bottom of the cassette Buffer Core Gel Tension cam locked Lower anode Buffer Chamber Gap used to fill Lower anode Buffer Chamber Y with Running Buffer Front of XCell SureLock Mini Cell Rear of XCell SureLock Mini Cell Align the lid on the Buffer Core The lid can only be firmly seated if the electrode is aligned over the banana plug on the right If the lid is not properly seated no power will go through the mini cell With the power off connect the electrode cords to power supply red to jack black to jack Caution Power must be off before connecting the XCell SureLock Mini Cell to the power supply Turn on the power See page 13 for running conditions for different gel types Disassembling the XCell SureLock Mini Cell Procedure 10 At the end of the run turn off the power and disconnect the cables from the power supply Remove the lid and unlock the Gel Tension Lever There is no need to remove the Gel Tension Wedge Remove the gel cassettes from the mini cell Handle gel cassettes by their edges only 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 o
21. ssette Slot faces out zi Side View 7 Ifyou are running only one gel replace the rear gel cassette with the Buffer Dam Continued on next page Assembling the XCell SureLock Mini Cell Continued Procedure 8 Pull forward on the Gel Tension Lever in a continued direction towards the front of the XCell SureLock unit until lever comes to a firm stop and the gels or gel buffer dam appear snug against the buffer core see figure below Unlocked Position Locked Position Buffer Core Buffer Core 1 Lower anode Gel Tension Wedge Lower anode J Gel Tension Wedge Buffer Chamber unlocked Buffer Chamber locked t x x o O Front Front R Gel 1 X d Gel 1 Gel When fully assembled cassettes and Buffer Core are in place and Gel Tension Wedge is locked into position Review the information provided on page 12 to troubleshoot your experiments Gel Electrophoresis Protocol Sample 1 Prepare your samples using the appropriate Loading sample buffer at the desired protein concentration 2 Prepare the appropriate running buffer see page 13 3 The Upper Buffer Chamber cathode is the void formed between the two gel cassettes or one cassette and the buffer dam on each side of the buffer core 4 Fillthe Upper Buffer Chamber with 200 mL of
22. tive Running Buffer 10X 500 mL LC2672 Novex Tris Glycine Native Sample Buffer 2X 20mL LC2673 Novex Tris Glycine Transfer Buffer 25X 500 mL LC3675 Novex Tricine SDS Running Buffer 10X 500 mL LC1675 Novex Tricine SDS Sample Buffer 2X 20mL LC1676 Novex IEF Cathode Buffer pH 3 10 10X 125 mL LC5310 Novex IEF Cathode Buffer pH 3 7 10X 125 mL LC5370 Novex IEF Anode Buffer for both pH ranges 50X 100 mL LC5300 Novex IEF Sample Buffer pH 3 10 2X 25mL LC5311 Novex IEF Sample Buffer pH 3 7 2X 25mL LC5371 Novex TBE Running Buffer 5X 1L LC6675 Novex Hi Density TBE Sample Buffer 5X 10 mL LC6678 Novex TBE Urea Sample Buffer 2X 10 mL LC6876 Novex Prep TBE Urea Sample Buffer 2X 20mL LC6877 Novex Zymogram Renaturing Buffer 10X 500 mL LC2670 Novex Zymogram Developing Buffer 10X 500 mL LC2671 16 Technical Support World Wide Visit the Invitrogen website at www invitrogen com for Web 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 Invitrogen 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 web page www invitrogen com Corporate Headquarters 5791 Van Allen Way Carlsbad CA 92008 USA T
23. uffer LC2673 LC2672 Expected Current Start 6 12 mA End 3 6 mA NuPAGE NuPAGE LDS NuPAGE MES SDS Voltage 35 50 Bis Tris SDS Sample Buffer Running Buffer 200V Constant minutes PAGE NP0007 NP0002 or Expected Current i deriaturing NuPAGE MOPS Start 110 125 mA non SDS Running Buffer End 70 80 mA reducing NP0001 RN NuPAGE Add NuPAGE Add NuPAGE Bis Tris SDS Sample Antioxidant PAGE Reducing Agent NP0005 to denaturing NP0004 to running buffer reducing sample buffer NuPAGE NuPAGE LDS NuPAGE Tris Voltage 50 minutes Tris Acetate Sample Buffer Acetate Running 150V Constant SDS PAGE NP0007 Buffer LA0041 Expected Current denaturing Start 40 55 mA NuPAGE Add NuPAGE Add NuPAGE End 25 40 mA Tris Acetate Sample Antioxidant SDS PAGE Reducing Agent NP0005 to denaturing NP0004 to running buffer reducing sample buffer Current readings are per gel Continued on next page 13 Buffers and Running Conditions Continued Gel Type Sample Running Buffer Power Settings Run Buffer Time Tricine SDS Tricine SDS Tricine SDS Voltage 90 minutes PAGE Sample Buffer Running Buffer 125V Constant denaturing LC1676 LC1675 Expected Current non reducing Start 80 mA End 40 mA Zymogram Tris Glycine Tris Glycine SDS Voltage 90 minutes SDS Sample Running Buffer 125V Constant Buffer LC2676 LC2675 Expected Current St
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