User Manual
Contents
1. 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 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 warranty will be in lieu of all warranties of fitness and in lieu of the warranty of merchantability 24 GenScript The Biology CRO GenScript USA Inc 120 Centennial Ave Piscataway NJ 08854 Tel 732 885 9188 732 885 9688 Fax 732 210 0262 732 885 5878 Email estain genscript com2. I 1 1 1 L 1 1 CBB 1 I if 1 1 F uf O Graphite Anode 40H 4e 0 2H O Features Important features of the eStain Protein Staining System are listed below Unique electric staining technology created for fast reliable and efficient protein gel staining in as few as 7 minutes User friendly electric staining unit allowing for easy and convenient procedures Consumable staining pads offering convenience without the need for additional solutions and buffers High staining efficiency as conventional Coomassie blue staining method Compatible for use with various types of mini polyacrylamide gels 10 eStain Protein Staining System www genscript com Quick Reference Guide Introduction A quick reference guide for operating the eStain Protein Staining Device is provided below Mode Action Sound Light Display eStain plugged in Connect eStain Steady upper Default Device to an electrical light running outlet and power time 000 switch is on eStain device and Place staining pads Steady upper Default staining pad assembled on the device and close light running lid time 000 Time selection Press Time buttonto Steady upper Running select desired running light time time defined Run Press Start Stop flashing upper Count down button light time End of run Automatic Continuous beeping Steady upper Default running for 2 minutes light time 000 Checking using freque
3. 250 1xeStain Anode Pad each pack eStain Protein Staining Pad G 250 contains 1xeStain Cathode Pad G 250 1xeStain Anode Pad The following eStain Graphite Electrode is available from GenScript Product Cat No eStain Graphite Electrode 11mm 1 pak L02014 staining replace it with a new one For best results when a Graphite Electrode was used for 200 times of electric eStain Protein Staining System www genscript com Product Specifications eStain Protein Staining Device Specifications eStain Protein Staining Pads Specifications Weight Dimensions Electrical Parameters Built in Features Compatibility Materials Operating Temperature Forceps Shovel 2 3 kg 200 mm I x180 mm w x90 mm h 100 120 V 220 240 V 50 60 Hz 3 2A Digital display alarm light LED Suitable for electric staining of proteins in mini polyacrylamide gels Acrylonitrile Butadiene Styrene Polycarbonate Aluminum Titanium Plasticized silicone 15 40 C Stainless steel 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 Protein Staining Device The specifications of the eStain Protein Staining Pads are listed below eStain Cathode Pad R 250 or G 250 size Materials eStain Anode Pad size M
4. Technical Support or visit www genscript com eStain Protein Staining System www genscript com Introduction System Overview The eStain Protein Staining System is a unique electric staining system that allows you to quickly reliably and efficiently stain proteins in various types of mini polyacrylamide gels with Coomassie blue dye reagents It consists of the eStain Protein Staining Device and eStain Protein Staining Pads The proprietary electric staining technology of the eStain Protein Staining Device combined with the eStain Protein Staining Pads applies a high electric force generated between graphite anode and titanium cathode to allow for quick and directional movement of negatively charged Coomassie blue dye reagents into the gel matrix to stain the proteins and also the quick and directional movement of the free staining reagents out of the gel matrix to destain the gel matrix within only 7 minutes or less The eStain Protein Staining System integrates three steps of conventional Coomassie blue staining method into a single step and greatly cuts down the time required for protein staining analysis The proteins stained using the eStain Protein Staining System exhibit as high detection sensitivity as proteins stained using conventional Coomassie blue staining method System Components The eStain Protein Staining System consists of eStain Protein Staining Device The eStain Protein Staining Devic
5. a washing step by soaking the Graphite Electrode in distilled water for half an hour and then drying it with a dry cloth or paper towel 20 eStain Protein Staining System www genscript com Troubleshooting Introduction Review the information below to troubleshoot your experiments using the eStain Protein Staining Device and eStain Protein Staining Pad Problem Cause Solution The upper status light doesn t flash during electric staining process Incomplete electric circuit due to improper assembly of the staining pads Ensure the staining stack is assembled correctly use the eStain Anode Pad first followed by the gel Gel Window and eStain Cathode Pad The upper and lower status lights flash simultaneously Excessive current is flowing through the Device Check the staining stack and ensure Gel Window covered correctly on the gel The stained gel has very tinged or nearly invisible protein bands with high blue background 1 The staining time is not long enough 2 Cooling the electrodes for too long a time 3 The graphite electrode has been used for 200 times or above 1 Cover Gel Window and the eStain Cathode Pad again and perform electric staining for 2 more minutes 2 Follow the instruction to perform cooling procedure 3 Replace the worn graphite electrode using a new one The stained gel has very tinged or nearly invisi
6. the eStain Anode Pad Gently remove air bubbles between gel and anode pad using the small shovel supplied with the device 14 eStain Protein Staining System www genscript com Protocols continued Assembling Staining 6 Select appropriate Gel Window according to actual size of the gel see the Stack with eStain Pads table below Place Gel Window on the gel Ensure that the Gel Window fully and Pre run Gel cover the margin of the anode pad continued Pre run gel Gel Window Gel with size 66 x 56mm to 80 x 60 mm Gel Windows 76 x 64mm Gel with size lt 66 x 56 mm Gel Windows 66 x 56mm Note Gel Window is used as the spacer between anode pad and cathode pad to prevent short circuit The edges of the pre run gel may also be covered by Gel Window and will probably be stained to deep blue after running an electric staining program 7 Tear the sealing of the 1 x eStain Cathode Pad package Remove the eStain Cathode Pad from the package and place it on the gel 8 Pull back the handle bar to close the lid of eStain Device while keeping the bar vertical to the lid and then return the bar to horizontal position 15 eStain Protein Staining System www genscript com Protocols continued Performing electric After assembling the staining stack perform electric staining as described staining below 1 Press the Time button to set appropriate running time based on the gel thicknes
7. the eStain Protein Staining Device works at staining mode press and hold Time button for 2 seconds to toggle to numbering mode If the Digital Display shows 200 or a number greater than 200 perform the replacing protocol as describe below 2 Switch Off the eStain Protein Staining Device and then open the lid of the device 3 Use small shovel to pry out the worn Graphite Electrode along the gap at either side of the anode tank afe eStain Protein Staining System www genscript com Protocols continued Replacing the eStain Graphite Electrode continued 4 Tear the sealing of a new eStain Graphite Electrode package and take the new Graphite Electrode out of the package Place the new Graphite Electrode into the anode tank and close the lid of the device 6 GenScript orn tee Dos Coenen 2m he toro 5 Switch On the eStain Device Press and hold Time button for 2 seconds to toggle to numbering mode 249 eStain Protein Staining System www genscript com Protocols continued Replacing the eStain Graphite Electrode continued 6 Press Reset button to zero the staining times After successfully installing Graphite Electrode into the eStain Device you are ready to use the eStain Device for staining applications again Note For the best results after using Graphite Electrode per 40 times of electric staining you may also need to perform
8. GenScript The Biology CRO e Stain Protein Staining System For electric staining of proteins in mini polyacrylamide gels Version 2 0 June 4 2010 User Manual eStain Protein Staining System www genscript com Table of Contents Product Contents 223 s22ssevseess so esoo nose soe sues sr ogaee ue a a a 1 Product Specifications s ssti4 ssc ape e a Soe 3 Stain Protein Staining Device lt 22 lt 9 s gt e ssseecs stesesecesesetesseeeussrepessssceess 5 Accessory Products a tt een enn ene ene en enn nee 7 Introduction a 8 Quick Relerence Guide lt ste2e ss2css2oncs gt trees 11 PIOlOCOlS em eae ee ater a enn eae Pe eee es aoe gee sts oe see eee 12 Troubleshooting 21 Examples ot Results gt AE 22 o weeclages RERESET ARER HA 23 Warranty a 24 eStain Protein Staining System www genscript com Product Contents Type of products eStain Protein Staining Device Contents Upon Receiving the Instrument eStain Protein Staining Pads This manual is supplied with the following products Product Cat No eStain Protein Staining Device L02010 The contents of the eStain Protein Staining Device are listed below Component Quantity eStain Protein Staining Device 1 each Specific Power Cord based on the type of unit o
9. MO900 Samples of 5ul New England Biolabs Protein Ladder P7703S in each lane were separated on a 4 20 Bis Tris gel and stained using eStain Protein Staining System R 250 e ve eStain Protein Staining System www genscript com Technical Support Web Resources Contact Us 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 product information and special offers For more information or technical assistance call write fax or email GenScript USA Inc 120 Centennial Ave Piscataway NJ 08854 Tel 732 885 9188 732 885 9688 Fax 732 210 0262 732 885 5878 Email estain genscript com DB eStain Protein Staining System www genscript com Warranty eStain Protein Staining Device GenScript warrants that eStain 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
10. aterials Gel Window Inter frame size Materials 8cm l x 7cm w x2 5mm thick Blotting filter paper presoaked with proprietary cathode buffer containing CBB R 250 or G 250 8cm I x 7cm w x2 5mm thick Blotting filter paper presoaked with proprietary anode buffer 66mmx56mm 76mmx64mm Polycarbonate eStain Protein Staining System www genscript com Product Specifications continued eStain Graphite Electrode Specifications The eStain Graphite Electrode is used as the replaceable anode of eStain Protein Staining Device and available separately from GenScript The specifications for eStain Graphite Electrode are listed below eStain Graphite Electrode 11mm Dimensions 100 mm I x90 mm w x11mm h Weight 180g Materials Powdered carbon clay eStain Protein Staining System www genscript com eStain Protein Staining Device Front View of The front top view showing various parts of the eStain Protein Staining Device eStain Device is shown below Control Panel Handle Bar Rear View of The rear view showing various parts of the eStain Protein Staining Device eStain Device is shown below Power Switch AC Inlet eStain Protein Staining System www genscript com eStain Protein Staining Device continued Control Panel of The control panel of the eStain Protein Staining Device is described below eStain Device When upper status lig
11. ble protein bands with clear background The staining time is too long 2 The electrodes have not been cooled or the cooling time is not long enough 1 Re assemble the staining stack using a new set of eStain Protein Staining Pad and then perform another 7 min electric staining 2 Follow the instruction to perform cooling procedure The stained gel has hazy blue spots 1 The gel has not been destained sufficiently 2 Cooling the electrodes for too long a time 3 Air bubbles present in staining stack interfering gel staining 1 Cover Gel Window and the eStain Cathode Pad again and perform electric staining for 1 more minute 2 Follow the instruction to perform cooling procedure 3 Be sure to remove all air bubbles present in the staining stack by using the small shovel supplied with eStain Device S94 eStain Protein Staining System www genscript com Examples of Results Tris Glycine gel results Bis Tris gel results Samples were separated on a 12 Tris Glycine gel and stained using eStain Protein Staining System R 250 123 4 5 6 7 8 Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 Cr cern Lane 7 Lane 8 500ng rabbit IgG 10ul E coli lysate 200ng reduced BSA 125ng reduced BSA 50ng reduced BSA 15ng reduced BSA 10ul New England Biolabs Protein Ladder Cat No P7703S 10ul GenScript Smart Middle Range Protein Standard Cat No M
12. e 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 eStain Protein Staining Pads are disposable pads to perform electric staining of proteins in gels with Coomassie blue dye Two types of eStain Protein Staining Pad R 250 and G 250 are available separately from GenScript and for different requirements Each pack 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 any solutions and buffers eStain Protein Staining System www genscript com Introduction continued System Components eStain Graphite Electrode continued The eStain Graphite Electrode serves as the replaceable anode of the eStain Protein Staining Device For the common use one eStain Graphite Electrode 11mm is pre installed inside the eStain Protein Staining Device to constitute a complete electric staining unit Each eStain Graphite Electrode is produced with special process and can be steadily used for 200 times of electric staining System Mechanism The eStain Protein Staining System is based on the electric staining concept which utilizes the unique and pa
13. fully separate the stained gel from the staining stack and proceed to be photographed 3 Discard the used eStain Protein Staining Pad Note Do not re use the eStain Protein Staining Pad after staining Discard after each use 4 Clean the titanium cathode plate graphite anode plate and its surrounding area with a dry cloth or paper towel Cooling Graphite For continuous use and best results it is necessary to cool down both graphite Anode and Titanium anode and titanium cathode to room temperature before the next run Place the Cathode pre frozen ice pack on the graphite anode close the lid gently to let the titanium cathode also touches the ice pack Do not force to close the lid otherwise the ice pack will damage the device At this point the eStain Protein Staining Device is ready for another run If you are not using the device turn Off the power switch located on the back of the device st eStain Protein Staining System www genscript com Protocols continued Replacing the eStain Graphite Electrode During electric staining process the eStain Graphite Electrode will absorb ions from anode pad and as well as lose carbon composition thereby changing the characteristics of the Graphite Electrode and composition of the anode buffer For the best staining results after having been used for 200 times of electric staining the worn Graphite Electrode should be replaced by a new one 1 If
14. ht is on which indicates staining mode the Digital Display shows running time otherwise when both upper and lower status lights are on which indicates numbering mode the Digital Display shows how many times of electric staining the eStain Graphite Electrode has been used for The Reset button is used to clear parameters The Time button is used to set running time and toggle between staining mode and numbering mode The Start Stop button is used to activate stop the staining program Inside View of The inside view showing various parts of the eStain Protein Staining ioe F eStain Device Device is shown below Titanium Cathode Graphite Anode eStain Protein Staining System www genscript com Accessory Products eStain Protein Staining Pads eStain Graphite Electrode Additional Products eStain Protein Staining Pads are available separately from GenScript Ordering information is provided below Product Quantity Cat No eStain Protein Staining Pads 1 box L02011 R 250 20 pak eStain Protein Staining Pads 1 box L02012 G 250 20 pak eStain Graphite Electrode is available separately from GenScript Ordering information is provided below Product Quantity Cat No eStain Graphite Electrode 11mm 1 pak 1 unit L02014 The precast Express PAGE Gels as well as premade buffers or buffer powders are available from GenScript For details contact
15. ncy of graphite anode Press and hold Time button for 2 seconds Steady upper and lower light Staining times the graphite anode has been used for Replacement of worn graphite anode Switch off the device and replace the worn graphite anode with a new one sfs eStain Protein Staining System www genscript com Protocols Recommendations Installing the eStain Device To obtain the best results follow these recommendations Wear gloves at all times during the entire staining procedures to prevent contamination of pads and gels Avoid using expired eStain Protein Staining Pads Always use the pads before the specified expiration date printed on the package Always keep the Ice Pack supplied with eStain Device stored at 20 C or below to facilitate cooling electrodes procedure Check the Power Cord supplied with the unit to ensure that the cord is compatible with the local socket format Place the eStain Protein Staining Device on a level laboratory bench Keep the area around the device clear to ensure proper ventilation of the unit 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 Ensure the AC power switch is in the Off position Attach the power cord to the AC inlet and then to the electrical outlet Use only properly grounded AC outlets and power cords When the electropho
16. rdered for U S Canada China 1 each Taiwan Japan Europe or UK Forceps 1 each Shovel 1 each eStain Graphite Electrode 11mm 1 each installed inside the device Ice Pack 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 ordered 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 Staining Pads before the expiration date printed on the package eStain Protein Staining System www genscript com Product Contents continued eStain Protein Staining Pads continued eStain Graphite Electrode Component Quantity The eStain Protein Staining Pads R 250 20 pak contains eStain Protein Staining Pad R 250 Gel Window 66mmx56mm Gel Window 76mmx64mm 20 The eStain Protein Staining Pads G 250 20 pak contains eStain Protein Staining Pad G 250 Gel Window 66mmx56mm Gel Window 76mmx64mm 20 each pack eStain Protein Staining Pad R 250 contains 1xeStain Cathode Pad R
17. resis of your samples is almost complete press the power switch located on the rear of the device to turn ON the eStain Protein Staining Device The upper status light is on indicating you are using staining mode Digital Display shows the default parameters 000 You are ready to use the eStain Protein Staining Device for staining applications 12 eStain Protein Staining System www genscript com Protocols continued Assembling Staining 1 Lift up the handle bar to vertical position then push it forward to open the Stack with eStain lid of eStain Device Pads and Pre run Gel Stain 2 Remove one package labeled as eStain Protein Staining Pad from the eStain Protein Staining Pads box and tear the laminated sealing of the package Remove the two small packages respectively labeled as 1 x eStain Cathode Pad R 250 or G 250 and 1 x eStain Anode Pad 219 eStain Protein Staining System www genscript com Protocols continued Assembling Staining Stack with eStain Pads and Pre run Gel 3 Tear the sealing of the 1 x eStain Anode Pad package Remove the eStain Anode Pad from the package and place it on the anode plate of the eStain Protein Staining Device continued 4 Carefully remove the pre run gel containing your protein samples from the gel cassette and rinse the gel with 50 to 100 ml distilled water for about 1 minute 5 Place the gel on
18. s see table below If an undesired running time is set by mistake press Reset button to clear the wrong time and then press again the Time button to choose the desired running time Gel Thickness Running Time 0 75 mm 6 minutes 1 0mm 7 minutes 1 5mm 9 minutes Note During electric staining process the graphite electrode may be heated to 60 70 C For continuous use and best results it is necessary to cool down both graphite anode and titanium cathode to room temperature before the next run see next page for cooling procedures 2 Press the Start Stop button to activate the electric staining program The running time begins to count down and upper status light keeps flashing during the whole staining program eStain Protein Staining System www genscript com Protocols continued Performing electric staining continued 3 Atthe end of the staining current automatically shuts off and the eStain Protein Staining Device signals the end of staining with repeated beeping sounds Upper status light stops flashing and the Digital Display shows text 000 4 Press any button on the control panel to stop the beeping 5 Proceed to disassemble the stack and clean the device Disassembling and To obtain good staining results disassemble the staining stack right away after cleaning the ending the staining procedure TM eStain Device 1 Open the lid of eStain Protein Staining Device 2 Care
19. tent pending technology developed by GenScript To use the eStain Protein Staining System for in gel protein staining assemble the eStain Protein Staining Pad with your pre run gel on the eStain Protein Staining Device Similar to semi dry blotting the eStain Cathode Pad and eStain Anode Pad act as ion reservoirs that contain the appropriate anode and cathode buffers incorporated into the gel matrix Simultaneously the eStain Cathode Pad presoaked with proprietary cathode buffer containing CBB dye R 250 or G250 also supplies negatively charged staining reagents The high electric force generated between graphite anode and titanium cathode allows for rapid and directional movement of negatively charged Coomassie blue dye reagents into the gel matrix to stain the proteins and also the homogenous movement of the free staining reagents out of the gel matrix to destain the gel matrix within only 7 minutes or less Therefore the eStain Protein Staining System provides fast convenient and reliable in gel protein staining without the need to prepare any solutions and buffers eStain Protein Staining System www genscript com Introduction continued System Mechanism Schematic mechanism of eStain Protein Staining System showing the continued movement of Coomassie blue dye Titanium Cathode E 2e 2H 0 gt H 20H eStain Cathode Pad CBB dye Pre run gel dye eStain Anode Pad 1 1
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