IPGbox IPGbox Kit - GE Healthcare Life Sciences
Contents
1. Immobiline DryStrip gel length cm 7 11 13 18 24 Total volume per strip pl 125 200 250 340 450 1 Including sample if applied Instructions 28 9357 71 AA 5 Carefully pull off the cover film from Immobiline DryStrip gel and place Immobiline DryStrip gel into the slot gel side down Distribute the rehydration solution evenly under the strip Gently lift and lower the strip and slide it back and forth along the surface of the solution to get complete and even wetting of the entire gel Note Ensure that no bubbles are trapped under the Immobiline DryStrip gel Fig 3 Example of IPG strips in a Reswell Tray Gently close the lid of the IPGbox and allow the Immobiline DryStrip gels to rehydrate at room temperature for 10 24 hours Overnight is recommended Use the Immobiline DryStrip gels within 20 minutes of opening the IPGbox Discard the used Reswell Tray Note Do not store the Reswell Tray in IPGbox Long time pressure of a Reswell Tray can reduce the life span of the IPGbox Insert 4 Selection of rehydration solution Depending on sample and strip select the appropriate rehydration solution Three options are available DeStreak Rehydration Solution 17 6003 19 with DeStreak reagent urea thiourea and CHAPS 2x Sample buffer rehydration solution stock 1 with urea thiourea and CHAPS recipe see Section 5 2x Sample buffer rehydration solution stock 2 with urea and CHAPS recipe see Section 5 DeStreak2. being used 1 If IPGphor standard Strip Holder or Immobiline OryStrip 7 11NL or 3 11NL is used use 0 5 IPG Buffer instead of 2 IPG Buffer as final concentration Rehydration solution made from stock Reagent Quantity Final concentration 2x Sample buffer rehydration solution stock 1or2 2 5 ml IPG Buffer same pH interval as the Immobiline 50 ult 2 DryStrip gels being used DTT 7 mg 1 If IPGphor standard Strip Holder or Immobiline DryStrip 7 11NL or 3 11NL is used use 0 5 IPG Buffer instead of 2 IPG Buffer as final concentration Instructions 28 9357 71 AA 6 Ordering information Product Pack size Code No IPGbox complete 1 IPGbox 1 IPGbox Kit 28 9334 65 IPGbox Kit 10 Reswell Trays 1 IPGbox Insert 28 9334 92 Related product Pack size Code No DeStreak Rehydration Solution 5x3ml 17 6003 19 Urea 500g 17 1319 01 Thiourea 100g RPN6301 CHAPS 1g 17 1314 01 Ettan IPGphor 3 Isoelectric Focusing Unit 1 11 0033 64 IPGphor Manifold Ceramic Tray 1 80 6498 57 Electrode set 1 80 6498 76 Sample cups 120 80 6498 95 PGphor Strip Holder Cleaning Solution 950 ml 80 6452 78 PG Buffer pH 3 5 5 0 1ml 17 6002 02 PG Buffer pH 4 5 5 5 1ml 17 6002 04 PG Buffer pH 5 0 6 0 1ml 17 6002 05 PG Buffer pH 5 5 6 7 1ml 17 6002 06 PG Buffer pH 4 7 1ml 17 6000 86 PG Buffer pH 6 11 1ml 17 6001 78 PG Buffer pH 7 11 NL Non Linear 1ml 17 6004 39 PG Buffer pH 3 10 NL 1ml 17 600
3. the Immobiline Drystrip gels are ready to use The IPGbox Insert keeps the conditions optimal for rehydration and is positioned inside the lid of IPGbox It can be used in ten reswelling cycles before disposal Immobiline DryStrip Cover Fluid is no longer required The rehydration solution is selected to fit the research experiment There are three options one with DeStreak reagent the DeStreak Rehydration Solution and two that you mix yourself The pH is adjusted by addition of desired IPG Buffer IPGbox Reswell Tray Fig 2 IPGbox parts 2 Instructions 28 9357 71 AA 2 Maintenance Assemble IPGbox and change IPGbox Insert To assemble IPGbox for the first time start at step 3 To change IPGbox Insert follow all steps IPGbox Insert should be changed for every 10th rehydration Note Do not touch the flat surface of the IPGbox Insert It is important to keep the flat surface clean and free from heavy pressure Note A convenient way to keep track of the number of rehydrations is to change IPGbox Insert every time a new package 10 Reswell Trays 1 IPGbox Insert is opened 2 Pull out the old IPGbox Insert from the lid by pulling in the plastic corners of the IPGbox Insert Discard the old IPGbox Insert 3 Take out a new IPGbox Insert from its protective plastic bag Hold only on the plastic edges to keep the flat surface intact 4 Carefully push on the edges of the IPGbox Insert to snap the IPGbox Insert in
4. 0 88 PG Buffer pH 3 10 1ml 17 6000 87 PG Buffer pH 3 11 NL 1ml 17 6004 40 Related product 7 cm 11 cm 13 cm 18 cm 24 cm mmobiline DryStrip pH 3 10 NL 17 6001 12 17 6001 15 7 1235 01 7 6002 45 mmobiline DryStrip pH 3 10 17 6001 11 8 1016 61 17 6001 14 7 1234 01 7 6002 44 mmobiline DryStrip pH 3 11 NL 17 6003 73 7 6003 74 17 6003 75 7 6003 76 7 6003 77 mmobiline DryStrip pH 3 7 NL 7 6002 43 mmobiline DryStrip pH 4 7 17 6001 10 8 1016 60 17 6001 13 7 1233 01 7 6002 46 mmobiline DryStrip pH 6 9 7 6001 88 7 6002 47 mmobiline DryStrip pH 6 11 17 6001 94 7 6001 95 17 6001 96 7 6001 97 mmobiline DryStrip pH 3 5 6 NL 17 6003 53 7 6003 54 17 6003 55 7 6003 56 7 6003 57 mmobiline DryStrip pH 5 3 6 5 17 6003 58 7 6003 59 17 6003 60 7 6003 61 7 6003 62 mmobiline DryStrip pH 6 2 7 5 17 6003 63 7 6003 64 17 6003 65 7 6003 66 7 6003 67 mmobiline DryStrip pH 7 11 NL 17 6003 68 7 6003 69 17 6003 70 7 6003 71 7 6003 72 nstructions 28 9357 71 AA Related literature Code No 2 D Electrophoresis Principles and Methods Appendix Ettan DIGE System User Manual 80 6429 60 18 1173 17 For contact information for your local office please visit www gelifesciences com contact GE Healthcare Bio Sciences AB Bj rkgatan 30 751 84 Uppsala Sweden www gelifesciences com 2D imagination at work GE imagination at work and GE monogram are trademarks of General Electric Company IPGbox Immobiline and DeStreak are trademarks of GE Hea
5. GE Healthcare Instructions 28 9357 71 AA IPGbox IPGbox IPGbox Kit IPGbox is used together with the IPGbox Kit to rehydrate Immobiline DryStrip gels in a convenient way The IPGbox Kit contains Reswell Trays and an IPGbox Insert both disposable This instruction contains information about how to Assemble IPGbox Clean IPGbox Change IPGbox Insert Select rehydration solution Prepare rehydration solution Rehydrate Immobiline Drystrip gels Fig 1 IPGbox Intended use The IPGbox Reswell Trays and IPGbox Insert are intended for research use only and shall not be used in any clinical or in vitro procedures for diagnostic purposes Table of contents aM a Luscoe Uo fo 0 perereeprerererertttetrescecereteeter tc S E OS 2 AMT GIN ST NG NCS Sigane ernn rE aE eE a E EE AEE E E e NEEE RENTENE EERE 3 3 Rehydratiom protol senson E R A NS 4 4 Selection of rehydration solution 0 eeesssesssssssssessesescccceccecesssssssssssssnssssssessecseceeceeeeesessesssssssssssanunueseees 5 5 Recipes GOrdering WAFOPM ATOM gnerra a e KERE EN A E REEE TE conse 7 1 Introduction Immobiline Drystrip gels require rehydration before use IPGbox is a convenient product for rehydration of Immobiline DryStrip gels A Reswell Tray is placed in the IPGbox rehydration solution is filled into the slots in the Reswell Tray and the Immobiline Drystrip gels are immersed into the rehydration solution After 10 hours or over night of reswelling
6. Rehydration Solution contains DeStreak reagent and optimized concentrations of urea thiourea and CHAPS DeStreak in the rehydration solution is recommended for strips with a pH above 7 If sample is added in the rehydration solution when performing In Gel rehydration DeStreak Rehydration Solution is not recommended The pH is adjusted by addition of IPG Buffer with same pH interval as the IPG strip used Detailed information on selection and making of rehydration solution can be found in the Appendix of the technical handbook 2 D Electrophoresis Principles and Methods 80 6429 60 or in Ettan DIGE System User Manual 18 1173 17 Instructions 28 9357 71 AA 5 Recipes Stock solutions 2x Sample buffer rehydration solution stock 1 Reagent Quantity Final concentration Urea M 60 06 10 5g 7M Thiourea M 76 12 3 89g 2M CHAPS M 614 89 0 5g 2 w v Distilled water Make up to 25 ml Small aliquots e g 2 5 ml can be stored at 20 C Stable for 6 months 2x Sample buffer rehydration solution stock 2 Reagent Quantity Final concentration Urea M 60 06 12g 8M CHAPS M 614 89 0 59 2 w v Distilled water Make up to 25 ml Small aliquots e g 2 5 ml can be stored at 20 C Stable for 6 months Working solutions DeStreak Rehydration Solution Reagent Quantity Final concentration DeStreak Rehydration Solution 17 6003 19 3 ml IPG Buffer same pH interval as the Immobiline 60 ult 2 DryStrip gels
7. lthcare companies All third party trademarks are the property of their respective owners 2008 General Electric Company All rights reserved All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them A copy of these terms and conditions is available on request Contact your local GE Healthcare representative for the most current information GE Healthcare UK Ltd Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare Bio Sciences KK Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan 28 9357 71 AA 04 2008
8. to the lid Instructions 28 9357 71 AA 3 5 Mount the lid by lowering the lid at 90 angle to the lower part Clean IPGbox Insert When necessary wipe the surface of IPGbox Insert carefully with a soft tissue moistened with 70 ethanol Clean IPGbox Clean IPGbox when necessary by 1 2 3 Open IPGbox and remove IPGbox Insert if present Handwash IPGbox with PGphor Strip Holder Cleaning Solution and rinse with water Check that IPGbox is dry before IPGbox Insert is mounted 3 Rehydration protocol Protocol for rehydration of Immobiline DryStrip gels using IPGbox and the Reswell Tray 1 2 3 Select and prepare a rehydration solution see Section 4 and 5 Change the IPGbox Insert if required see Section 2 Place the IPGbox on a levelled table and place a Reswell Tray in the IPGbox Ensure that the Reswell Tray and the IPGbox Insert are clean and dry If required wipe the IPGbox Insert with a soft tissue moistened with 70 ethanol Pipette the appropriate volume of selected rehydration solution and sample if applied evenly over the slots corresponding to the length of the DryStrip gels See the table below for rehydration solution volumes Strip lengths are measured from the straight ended part of the wells of the Reswell Tray and marked with lines on the top of the Reswell Tray Note For complete sample uptake do not apply excess rehydration solution Rehydration solution volume per Immobiline DryStrip gel
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