eStain user manual
Contents
1. 15 eStain 2 0 Protein Staining System Protocols continued Performing Electric Staining continued Disassembling and Cleaning the eStain 2 0 Device www genscript com 3 At the end of the staining current automatically shuts off and the eStain 2 0 Protein Staining Device signals the end of staining with repeated beeping sounds The right status light stops flashing and the lower four digits show text 00 00 4 Press any button on the control panel to stop the beeping 5 Proceed to disassemble the stack and clean the device To obtain consistent staining results disassemble the staining stack right away after ending the staining procedure 1 Open the closed lid by pressing the Open button 2 Carefully separate the stained gel from the staining stack and proceed to document the gel image 3 Discard the used eStain Protein Staining Pad Note Do Not reuse the eStain Protein Staining Pad Discard after each use 4 Take the eStain 2 0 Graphite Electrode out of the device dry the contact rods with a dry cloth or tissue paper Take care not to lose the spring on the rod located near the triangle symbols before placing back into the device Caution Do Not clean the eStain 2 0 Graphite Electrode immediately after ending the staining program since the graphite electrode is still hot after a run Wait for 3 to 5 minutes before proceeding to cleaning 5 Clean the titanium cathode and the anode tank with2. Schematic mechanism of eStain 2 0 Protein Staining System showing the Introduction continued movement of Coomassie blue dye is provided below Titanium Cathode 2e 2H O H 20H Mode Action Sound Light Display Sponge cushion and Insert the sponge Bar graphite electrode cushion and graphite i installed inside electrode in the eStain Cathode Pad eStain 2 0 Device anode tank eStain 2 0 Device Connect eStain 2 0 Steady right Default running plugged in Device to an light time 00 00 Pre run gel electrical outlet and power switch is on eStain Anode Pad eStain 2 0 Device Place staining stack Steady right Default running and staining stack on the device and light time 00 00 assembled close lid Graphite Anode Ct 40H gt O 2H O 4e Time selection Press Min and Sec Steady right Running time button to select light defined desired running time Run Press Start Stop flashing right Count down System Features Important features of the eStain 2 0 Protein Staining System are listed below button light time Innovative electric staining technology created for fast reliable and efficient protein gel staining in 7 minutes or less End of run Automatic Continuous beeping Steady right Default running User friendly electric staining unit allowing for easy and convenient for 2 minutes light time 00 00 procedures Consumable staining pads offering convenience without the need for Check
3. 2 0 Graphite Electrode into the eStain 2 0 Protein Staining Device you are ready to use the device for next staining application Note For best results after every 40 times of electric staining wash the eStain 2 0 Graphite Electrode by soaking in distilled water for half an hour followed by drying with tissue paper oe eStain 2 0 Protein Staining System Troubleshooting Problem The right status light doesn t flash during electric staining process The left and right status lights flash simultaneously The stained gel has very faint or nearly invisible protein bands with high blue background The stained gel has very faint or nearly invisible protein bands with clear background The stained gel has blue spots Blue spots observed at the bottom of the stained gel Cause Incomplete electric circuit due to improper assembly of the staining stack Excessive current is flowing through the eStain 2 0 Device 1 The staining time is not long enough 2 The eStain 2 0 Graphite Electrode has been used for more than 100 times The staining time is too long The gel has not been destained sufficiently 1 The spring is missing 2 The eStain 2 0 Graphite Electrode is installed in wrong direction 90 www genscript com Solution Ensure the staining stack is assembled correctly Use the eStain Anode Pad first followed by the gel and eStain Cathode Pad C
4. Device The specifications of the eStain Protein Staining Pads are listed below eStain Cathode Pad R 250 or G 250 size 90 mm lI x 80 mm w x 2 5 mm t Materials Blotting filter paper presoaked with proprietary cathode buffer containing CBB dye R 250 or G 250 eStain Anode Pad size 90 mm l x 80 mm w x 2 5 mm t Materials Blotting filter paper presoaked with proprietary anode buffer Absorbent Filter Paper Size 80 mm I x 70 mm w x 2 6 mm t Materials Vegetable fiber eStain 2 0 Protein Staining System www genscript com Product Specifications continued eStain Protein Staining Pads Specifications continued eStain 2 0 Graphite Electrode Specifications Sponge Cushion Size 104 mm I x 100 mm w x 5 mm t Materials Sponge The eStain 2 0 Graphite Electrode is used as the replaceable anode electrode of eStain 2 0 Protein Staining Device and available separately from GenScript The specifications for eStain 2 0 Graphite Electrode are listed below Dimensions 100 mm I x 92 mm w x 8 8 mm t Weight 206 g Materials Powdered carbon Clay Stainless steel eStain 2 0 Protein Staining System www genscript com eStain 2 0 Protein Staining Device Front View of eStain 2 0 Device Rear View of eStain 2 0 Device The front top view showing various parts of the eStain 2 0 Protein Staining Device is shown below OO Control Panel e Stalin 2 0
5. Open Button The rear view showing various parts of the eStain 2 0 Protein Staining Device is shown below AC Inlet Power Switch eStain 2 0 Protein Staining System www genscript com eStain 2 0 Protein Staining Device continued Control Panel of eStain 2 0 Device Top View of Openned eStain 2 0 Device The control panel of the eStain 2 0 Protein Staining Device is described below The Digital Display shows two rows of multi digits that specify the electric staining conditions as follows The upper three digits after text lt PN gt indicate number of usage of the eStain 2 0 Graphite Electrode The lower four digits specify the time of electric staining in minute and second The two status lights show the working modes of the eStain 2 0 Protein Staining Device When the right status light is on the device is switched on and working at staining mode when both left and right status lights are on the device is working at numbering mode The Reset button is used to clear parameters The Min button is used to shift between staining and numbering mode and to set running time Each short press will increase one minute Each long press 2 seconds will toggle working mode from staining to numbering or vice versa The Sec button is also used to set running time each press will increase 5 seconds The Start Stop button is used to activate stop the staining program The top view of openned eSta
6. Press the Open button then push back and close the lid of eStain 2 0 Protein Staining Device 4 Carefully remove the pre run gel containing your protein samples from the gel cassette and briefly rinse the gel with distilled water Re ee eStain 2 0 Protein Staining System Protocols continued Performing Electric Staining www genscript com After assembling the staining stack perform electric staining as described below Perform electric staining within 15 minutes after assembling the pads with the gel 1 Use the following parameters as starting points for electric staining based on gel thickness Gel Thickness Running Time 0 75 mm 6 7 minutes 1 0 mm 7 8 minutes 1 5mm 7 8 minutes Press the Min and Sec buttons to set appropriate running time If an undesired running time is set by mistake press Reset button to clear the wrong time and then press again the Min and Sec buttons to choose the desired running time g Stalin 2 0 i Note For best staining results it is recommended to use eStain 2 0 Protein Staining System at room temperature 22 28 C If the ambient temperature is below 22 C you may need to extend the running time to obtain satisfied staining results based on gel thickness 2 Press the Start Stop button to activate the electric staining program The running time begins to count down and right status light keeps flashing during the whole staining program
7. a dry cloth or tissue paper 6 Replace the used Sponge Cushions with new ones after 20 times of electric staining 7 Replace the used Absorbent Filter Paper in the waste tray with a new one after 20 times of electric staining At this point the eStain 2 0 Protein Staining Device is ready for another run If you are not using the device for a period of time turn off the power switch located on the back of the device take the graphite anode out and keep the lid open to let any solution left in the holes for the contact rods dry out For any other repairs and services contact Technical Support Do not perform any repairs or services on the eStain 2 0 Protein Staining Device by yourself to avoid damaging the device 16 eStain 2 0 Protein Staining System Protocols continued Replacing the eStain 2 0 Graphite Electrode www genscript com During electric staining process the eStain 2 0 Graphite Electrode will absorb ions from anode pad while losing carbon composition thereby changing the characteristics of the eStain 2 0 Graphite Electrode and composition of the anode buffer For best staining results after 100 uses of electric staining the worn eStain 2 0 Graphite Electrode should be replaced by a new one Caution Do Not replace the graphite electrode immediately after ending the staining program since the graphite electrode is still hot after a run Wait for 3 to 5 minutes before proceeding to the
8. new Absorbent Filter Paper is included in each box of f eStain Protein Staining Pads Installing the 1 Check the Power Cord supplied with the unit to ensure that the cord is af H eStain 2 0 Device compatible with local socket format 2 Place the eStain 2 0 Protein Staining Device on a leveled laboratory bench Keep the area around the device clear to ensure proper ventilation of the unit e Stain 2 0 3 For your safety Position the device properly such that the power switch and the AC inlet located on the rear of the unit are easily accessible 4 Ensure that the AC power switch is in the Off position 5 Open the closed lid of the eStain 2 0 Protein Staining Device by pressing 8 Attach the power cord to the AC inlet then to the electrical outlet the Open button Place one or two pieces of Sponge Cushion in the anode Use only properly grounded AC outlets and power cords tank depending on gel thickness For 1 5 mm gel use one piece of Sponge Cushion for 0 75 and 1 0 mm gel use two pieces of Sponge Cushion 9 When the electrophoresis of your samples is almost completed press the power switch located on the rear of the device to turn ON the eStain 2 0 Protein Staining Device The right status light is on indicating staining mode The lower four digits of the digital display show the default running time 00 00 Note After 20 times of electric staining replace the used Sponge Cushions with new ones A pai
9. replacement protocol 1 If the eStain 2 0 Protein Staining Device works at staining mode press and hold Min button for 2 seconds to toggle to numbering mode If the upper three digits show 100 or a number greater than 100 replace the eStain 2 0 Graphite Electrode as described below 2 Switch Off the eStain 2 0 Protein Staining Device Open the lid of the device and take the worn eStain 2 0 Graphite Electrode out of the device 3 Remove the new eStain 2 0 Graphite Electrode from the packing box Note Wear gloves when handling the eStain 2 0 Graphite Electrode SAP eStain 2 0 Protein Staining System Protocols continued Replacing the eStain 2 0 Graphite Electrode continued www genscript com 4 Insert the contact rods of the new eStain 2 0 Graphite Electrode into the holes in the anode tank Make sure that the triangle symbols at the edge of the graphite electrode face the Open button 5 Push the eStain 2 0 Graphite Electrode down gently until you hear a click 18 eStain 2 0 Protein Staining System Protocols continued Replacing the eStain 2 0 Graphite Electrode continued www genscript com 6 Close the lid and switch On the eStain 2 0 Protein Staining Device Press and hold Min button for 2 seconds to toggle to numbering mode When the upper three digits are flashing press Reset button to zero After successfully installing the eStain
10. Cushion when staining 1 5 mm gel use 2 pieces of Sponge Cushion when staining 0 75 and 1mm gel eStain 2 0 Protein Staining System www genscript com Examples of Results Tris Glycine Gel Samples were separated on a 12 Tris Glycine gel and stained using eStain 2 0 Results Protein Staining System with CBB R 250 1 2 3 4 5 6 7 8 9 10 Lane 1 1 ug mouse IgG mm Lane 2 200 ng recombinant His tagged protein MEER mi gt m Lane 3 10 ul E coli lysate TE Lane 4 5 ul E colilysate pas meet Lane 5 400 ng reduced BSA F lt lt Lane 100 ng reduced BSA ss ae Lane 7 50 ng reduced BSA Lane 8 20 ng reduced BSA Lane 9 10 ul New England Biolabs Protein Ladder Cat No P7703S Lane 10 5 ul New England Biolabs Protein Ladder Cat No P7703S Bis Tris Gel 7ul New England Biolabs Protein Ladder P7703S were loaded in each lane Results and separated on 4 20 Express PAGE Gel Bis Tris gel The gel was then stained using eStain 2 0 Protein Staining System with CBB R 250 sd eStain 2 0 Protein Staining System Technical Support Web Resources Contact Us www genscript com Visit the GenScript web site at www genscript com for 1 Technical resources including manuals vector maps and sequences application notes MSDSs FAQs formulations citations handbooks etc 2 Complete technical support contact information 3 Access to the GenScript Online Catalog 4 Additional produc
11. eStain 2 0 Protein Staining System For fast Goomassie blue staining of proteins in mini polyacrylamide gels Mar 28 2012 User Manual GenScript Transforming Biology Research GenScript USA Inc 860 Centennial Ave Piscataway NJ 08854 Tel 732 885 9188 732 885 9688 Fax 732 210 0262 732 885 5878 Email estain genscript com GenScript Transforming Biology Research eStain 2 0 Protein Staining System www genscript com Table of Contents a0 Ue 6 COMICAS aaa OB RE VERE Ee 1 Pioduct SPCCIICAllONS Mees oe EE ER eee eee eet 3 eStain 2 0 Protein Staining Device 5 Introduction 7 Quick Reference Guide EE EE EE EER EE EE EER ER EE EE EER ER EE n eens 10 gelees ER IE EE IE EIE a IE IE EE Ee 11 Troubles hoo lng sosio RA DE NE ED AE EE N RE eae 20 Examples Of RESUS Ee ER RD AE ORE EE DE EG SE ER ORE ER ere 22 Technical Support 7 7 7 7 93 Warranty 24 else eee eee 25 eStain 2 0 Protein Staining System Product Contents Type of Products eStain 2 0 Protein Staining Device Contents Upon Receiving the Instrument eStain Protein Staining Pads www genscript com This manual is supplied wi
12. heck if the staining stack is properly assembled and ensure full coverage of the gel 1 Cover the eStain Cathode Pad again and then perform electric staining for 1 or 2 more minutes 2 Replace the worn eStain 2 0 Graphite Electrode with a new one Do Not stain the gel using eStain 2 0 System again Instead stain the gel using traditional tri step method Invert the eStain Cathode Pad cover it back on top of the gel Perform electric staining for 1 or 2 more minutes 1 Fix the spring on the contact rod located close to triangle symbols 2 Follow the steps described in the protocol to install the eStain 2 0 Graphite Electrode in correct direction eStain 2 0 Protein Staining System Troubleshooting continued Problem Ring like spots observed on the protein bands It is hard to close the lid or the lid can t be closed tightly Cause 1 Air bubbles trapped in the assembled staining stack Small gel pieces attached on the surface of the gel The lid is not closed properly Extra Sponge Cushions are placed in the anode tank DA www genscript com Solution 1 Use the small shovel supplied with eStain 2 0 Device to press air bubbles out after staining stack assembly Make sure to remove all the gel pieces by washing the gel in distilled water prior to staining Follow the instruction on page 14 to properly close the lid Use 1 piece of Sponge
13. hich is or less Therefore the eStain 2 0 Protein Staining System provides fast comparable to the sensitivity of traditional method despite of their different working convenient and reliable in gel protein staining without the need to prepare nni additional buffers principles System Components The eStain 2 0 Protein Staining System consists of eStain 2 0 Protein Staining Device The eStain 2 0 Protein Staining Device is a user friendly electric staining unit that allows for fast convenient and efficient in gel protein staining with Coomassie blue dye reagents eStain Protein Staining Pads The disposable eStain Protein Staining Pads are used to perform electric staining of proteins in gels with Coomassie blue dye Two types of eStain Protein Staining Pads R 250 and G 250 are available separately from GenScript and for different requirements Each pack of eStain Protein Staining Pad contains an eStain Cathode Pad presoaked with proprietary cathode buffer containing CBB dye R 250 or G 250 and an eStain Anode Pad presoaked with proprietary anode buffer allowing for rapid convenient and reliable in gel protein staining without the need to prepare additional buffers eStain 2 0 Protein Staining System www genscript com eStain 2 0 Protein Staining System www genscript com Introduction continued Quick Reference Guide System Mechanism A quick reference guide for operating the eStain 2 0 Protein Staining Device
14. in 2 0 Protein Staining Device identifying various parts is shown below Titanium Cathode Graphite Anode eStain 2 0 Protein Staining System www genscript com eStain 2 0 Protein Staining System www genscript com Introduction Introduction continued types of mini polyacrylamide gels with Coomassie blue dye It consists of the eStain 2 0 Staining System for protein gel staining assemble the eStain Protein cathode to allow for quick and directional movement of negatively charged Coomassie Cathode Pad presoaked with cathode buffer containing CBB dye R 250 or blue dye into the gel matrix to stain the proteins and also the homogeneous movement of G250 supplies negatively charged staining reagents The voltage generated the unbound staining reagents out of the gel matrix to destain the gel matrix within only 7 between graphite anode and titanium cathode allows for rapid and directional minutes or less The eStain 2 0 Protein Staining System integrates three steps of movement of negatively charged Coomassie blue dye into the gel matrix to conventional Coomassie blue staining method into a single one and greatly cuts down the stain proteins as well as for the homogeneous movement of the unbound time required for protein staining analysis The eStain 2 0 Protein Staining System is able staining reagents out of the gel matrix to destain the gel within only 7 minutes to deliver high detection sensitivity down to several ng per protein band w
15. ing number of Press and hold Min Steady left Number of usage sage of graphite button for 2 seconds and right of the graphite additional buffers AAge ol grape u g grap anode lights electrode High staining efficiency as conventional Coomassie blue staining method Compatible for use with various types of mini polyacrylamide gels Replacement of worn graphite anode Switch off the device and replace the worn graphite anode with a new one lt 40 eStain 2 0 Protein Staining System www genscript com eStain 2 0 Protein Staining System www genscript com Protocols Protocols continued Recommendations Follow these recommendations for best results Installing the Note For a new eStain 2 0 Protein Staining Device two pieces of Sponge Cushion and 1 Wear gloves at all times during the entire staining procedures to prevent eStain 2 0 Device one unit of eStain 2 0 Graphite Electrode have already been installed inside the ti d unit for immediate use If staining 1 5 mm gel remove 1 piece of Soonge Cushion contamination of pads and gels continue 2 Avoid using expired eStain Protein Staining Pads Always use the pads 7 Pull out the waste tray from the right side of the device Place a new before the specified expiration date printed on the package Absorbent Filter Paper inside the tray and then push the tray back in Note After 20 times of electric staining replace the used Absorbent Filter Paper with a new one One piece of
16. lter Paper Sponge Cushion each pack of eStain Protein Staining Pad R 250 contains 1xeStain Cathode Pad R 250 1xeStain Anode Pad each pack of eStain Protein Staining Pad G 250 contains 1xeStain Cathode Pad G 250 1xeStain Anode Pad eStain 2 0 Graphite Electrode Product eStain 2 0 Graphite Electrode www genscript com Quantity 20 20 The following eStain 2 0 Graphite Electrode is available from GenScript Cat No L02017 For best results after 100 uses of electric staining replace the worn eStain 2 0 Graphite Electrode with a new one eStain 2 0 Protein Staining System Product Specifications eStain 2 0 Protein Staining Device Specifications eStain Protein Staining Pads Specifications www genscript com Weight 1 78 kg Dimensions 325 mm I x 195 mm w x 70 mm h Electrical Parameters 100 120 V 220 240 V 50 60 Hz 3 2 A Built in Features Digital display alarm light LED Compatibility Suitable for fast Coomassie blue staining of proteins in mini polyacrylamide gels Materials Acrylonitrile butadiene styrene Polycarbonate Aluminum Titanium Plasticized silicone Operating Temperature Room temperature Forceps Stainless steel Shovel Polycarbonate Tray Polycarbonate Avoid acetone dimethyl sulfoxide and acetic acid These reagents can erode or damage the device The eStain Protein Staining Pads are used with the eStain 2 0 Protein Staining
17. r of new Sponge Cushions are included in each box of eStain Protein Staining a You are ready to use the eStain 2 0 Protein Staining Device for staining applications 6 Insert the eStain 2 0 Graphite Electrode into the anode tank as described in Section Replacing the eStain 2 0 Graphite Electrode then close the lid of the device Tr 1E eStain 2 0 Protein Staining System www genscript com eStain 2 0 Protein Staining System www genscript com Protocols continued Protocols continued Assembling Staining 1 Open the closed lid by pressing the Open button Assembling Staining 5 Place the gel on the eStain Anode Pad Gently remove air bubbles Stack with eStain Stack with eStain between gel and anode pad using the small shovel supplied with the Pads and Pre run Gel Pads and Pre run Gel device continued 2 Remove one package labeled as eStain Protein Staining Pad from the eStain Protein Staining Pads box and tear the laminated sealing of the l 6 Tear the sealing of the 1xeStain Cathode Pad package Remove the package Remove the two small packages labeled 1xeStain Cathode Pad R 250 or G 250 and 1xeStain Anode Pad respectively eStain Cathode Pad from the package and place it on top of the gel 3 Tear the sealing of the 1xeStain Anode Pad package Remove the eStain Anode Pad from the package and place it on the anode plate of the eStain 2 0 Protein Staining Device 7
18. rranty will be in lieu of all warranties of fitness and in lieu of the warranty of merchantability 94 eStain 2 0 Protein Staining System www genscript com Appendix FEE TREAT E N ve Certificate No CECS 0020110523A VERIFICATION OF EMC COMPLIANCE INCIL DIRECTIVE 2004 108 EC nical Construction File And Manufacturi sesses For Above Mentioned Sample Safety Requirements of EU COUNCIL Saat Standards i 25 eStain 2 0 Protein Staining System www genscript com continued Appendix TE ni 1 a JNC L DIRECTIVE 2006 95 EC dIE Tr 105 0 President of CECS Abbe GE Westminster Grane Landaa SIF info cecsuk com 26 eStain 2 0 Protein Staining System www genscript com Appendix continued According to FCC Part 15B Certificate No SEM11058591 Responsible Party s Name NanJing Genscript Co Ltd Address No 78 Shuangbai Road Xuanwu District Nanjing China NanJing Genscript Co Ltd Address No 78 Shuangbai Road Xuanwu District Nanjing China Description of Product Multifunction Gel Processor Model No GS 01 GS 02 Trade Name eStain eBlot Report No STR11058051E 3 Compliance With Part 15B of FCC Rules z This device complies with Part 15 of the FCC Rules Operation is subject to f ad the following two conditions 1 This device may not cause harmful interference and 2 This device must accep
19. t any interference received including interference 4 Tested By ENG SEM Test Compliance Service Co Ltd 3 F Jinbao Commerce Building Xin an Fanshen Road andy S07 PS eManager d hel j Pi i a hy Er F The Certification of Verification shows that the tested sample technically compliances with the FCC Part 15 The certification applies to the tested sample above mentioned only and should not implied an assessment of the whole Tel 86 755 33663308 Fax 86 755 33663309 E mail ge 27
20. t information and special offers For more information or technical assistance call write fax or email GenScript USA Inc 860 Centennial Ave Piscataway NJ 08854 Tel 732 885 9188 732 885 9688 Fax 732 210 0262 732 885 5878 Email estain genscript com DA eStain 2 0 Protein Staining System Warranty eStain 2 0 Protein Staining Device www genscript com GenScript warrants that eStain 2 0 Protein Staining Device will be free from defects in material and workmanship for a period of one year from date of purchase If any defects occur in the product during this warranty period GenScript will at its option repair replace or refund the purchase price of this product at no charge to you The following defects however are specifically excluded 1 Defects caused by improper operation 2 Repair or modification done by anyone other than GenScript or an authorized agent 3 Use of fittings or other spare parts supplied by anyone other than GenScript 4 Damage caused by accident or misuse 5 Damage caused by disaster 6 Corrosion due to the use of improper solvent or sample For any inquiry or request for repair service contact GenScript after confirming the model and serial number of your instrument For your protection items being returned must be insured against possible damage or loss This warranty shall be limited to the replacement of defective products It is expressly agreed that this wa
21. th the following product Product Cat No eStain 2 0 Protein Staining Device L02016 The contents of the eStain 2 0 Protein Staining Device are listed below Component Quantity eStain 2 0 Protein Staining Device 1 each eStain 2 0 Graphite Electrode oe 1 each Installed inside the device Sponge Cushion ean 2 each Installed inside the device Regional Specific Power Cord 1 each Forceps 1 each Shovel 1 each Tray 1 each Examine the unit carefully for any damage incurred during transit File any damage claims with the carrier The warranty does not cover in transit damage The following eStain Protein Staining Pads are available from GenScript Product Cat No eStain Protein Staining Pads R 250 20 pak L02011 eStain Protein Staining Pads G 250 20 pak L02012 If you order the eStain Protein Staining Pads you will receive the components listed in the table below Store the eStain Protein Staining Pads at room temperature For best results use the eStain Protein Staining Pads before the expiration date printed on the package eStain 2 0 Protein Staining System Product Contents continued eStain Protein Staining Component Pads continued The eStain Protein Staining Pads R 250 20 pak contains eStain Protein Staining Pad R 250 Absorbent Filter Paper Sponge Cushion The eStain Protein Staining Pads G 250 20 pak contains eStain Protein Staining Pad G 250 Absorbent Fi
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