Cleaver Scientific CVS10CBS Instruction Manual Download Page 2

 

 

 

Contents 

Instruction Manual 

Catalogue Numbers 

Contents 

Safety Information 

Packing List 

Care and Maintenance 

Operating Instructions 

Usage Guidance and restrictions 

Setting up the Gel Tanks 

Protein Electrophoresis  

Casting Unit Preparation 

Gel Preparation 

Preparation of denatured protein samples for loading 

11 

Gel Pouring 

11 

 Gel Running 

14 

Protein Blotting  

16 

Setting up the blot sandwich 

16 

Blot Running Conditions 

17 

Solutions  

19 

References  

21 

Troubleshooting 

22 

Warranty 

                                                                                                                                   28

 

 

 

 

 

 

 

 

Summary of Contents for CVS10CBS

Page 1: ...gue Numbers CVS10CBS SB10 Record the following for your records Model _____________________ Catalogue No _____________________ Date of Delivery _____________________ Warranty Period __________________...

Page 2: ...e Guidance and restrictions 6 Setting up the Gel Tanks 6 Protein Electrophoresis 7 Casting Unit Preparation 7 Gel Preparation 9 Preparation of denatured protein samples for loading 11 Gel Pouring 11 G...

Page 3: ...tly in position The unit should not be used if there is any sign of damage to the external tank or lid Acrylamide is a powerful neurotoxin in solution form Polymerized gels can contain some unpolymeri...

Page 4: ...g List Checked by ________________________ Date ________________________ The packing lists should be referred to as soon as the units are received to ensure that all components have been included The...

Page 5: ...de dishwashing liquid Hexane and Aliphatic hydrocarbons The units should not be left to in detergents for more than 30 minutes The units should never come into contact with the following cleaning agen...

Page 6: ...ionally however a temporary conductivity caused by condensation must be expected Setting up the Gel Tanks Note Before setting up the Gel Tank please ensure that it has been properly cleaned and dried...

Page 7: ...ause leaks Gel Cassette Assembly Assemble the glass plates in the CVS10DRIM casting running insert so that the bottom of the glass plates and the spacers are perfectly aligned on a flat levelled surfa...

Page 8: ...When using the Slide Clamp Mini version simply slide both gates outwards until fully tightened When only one gel is being run the dummy plate must be used in the second position and fully tightened NO...

Page 9: ...e clamps or screws as preferred by the user For those that prefer to use the screws rather than clamps the screws can be simply inserted into the screw holes The clamps can be removed by placing each...

Page 10: ...olumes of solutions from the standard stock solutions These should be gently mixed avoiding generation of bubbles which will inhibit polymerization by removing free radicals Resolving Gel for 2 x 1mm...

Page 11: ...combine the protein sample and 4 X sample buffer It is always advisable to use protein markers in one of the end lanes to indicate sizes of bands These should be prepared according to the manufacture...

Page 12: ...n this use fresher stock solutions or add more APS and TEMED 7 Prepare the stacking gel solution 8 Before casting the stacking gel insert a piece of filter paper to dry the area in between the glass p...

Page 13: ...ideways with the end s of the tank and pressed into the recess Or these can be fitted down the front of the tank Note NEVER FIT THESE UNDERNEATH THE MODULE IN THE BOTTOM OF THE TANK AS THIS WILL PREVE...

Page 14: ...ve gel level Cooling packs are inserted behind the gels Outer Tank is filled to the maximum fill line Cooling is at a maximum 1000ml Gel Running 1 Fit the lid and connect to a power supply 2 Gels shou...

Page 15: ...door Twist key applying pressure to both the clamping door and the CVS10D side cheek The door will now click open Repeat this process until you have opened both the doors 4 Remove the glass plates The...

Page 16: ...and trapped bubble should be removed with a roller at each step during blotting sandwich assembly 1 Each blot sandwich should be set up according to the following order Cassette clamp ve black side pl...

Page 17: ...entation and order the blot sandwiches were loaded in This can be done by noting which samples were loaded adjacent to each electrode and by marking the membrane with a pencil on one corner 2 Use of a...

Page 18: ...he membrane is now ready to be probed Suggested Running Conditions please note that these are just guideline Duration of Blot SB10 One Hours Three Hours Overnight 16 hr 100V 125V 250mA 400mA 50V 200mA...

Page 19: ...o 200ml with Distilled Water Stock 4 X Stacking Tris 0 5 M Tris HCL pH6 8 0 4 SDS To 110ml Distilled Water add 12 12 g of Tris base Add 8ml of 10 SDS Adjust pH to 6 8 with 1N HCl Add Distilled Water t...

Page 20: ...03 g Bromophenol blue Aliquot into 1 5ml microcentrifuge tubes Store at 20 C Towbin Buffer 25mM Tris 192mM glycine 20 methanol pH8 3 Towbin Buffer SDS 25mM Tris 192mM glycine 20 methanol pH8 3 0 05 0...

Page 21: ...pH will vary according to the freshness of the reagents used References 1 Sambrook Fritsch and Maniatis Molecular Cloning A Laboratory Manual Second Edition Cold Spring Harbor Laboratory Press 1989 2...

Page 22: ...0 06 APS and 0 12 TEMED Gel does not polymerize Too little or too much APS or TEMED Failure to degas Temperature too low Poor quality acrylamide or bis Old APS Use 0 0 05 APS and 0 05 TEMED Degas mono...

Page 23: ...rument manual Poor staining sensitivity Dirty staining trays Insufficient stain volume Insufficient staining time Reuse of staining solution Clean staining trays and other equipment with laboratory gl...

Page 24: ...ing gel Uneven gel interface Remove salts from sample by dialysis or desalting column prior to sample preparation Use same buffer in samples as in gel Check buffer composition and dilution instruction...

Page 25: ...extent that it prevents binding to nitrocellulose Alternatively reduce methanol concentration and switch to PVDF Protein precipitating in gel Use SDS in transfer buffer SDS can increase transfer effi...

Page 26: ...brane PVDF Membrane is not completely wet Because of hydrophobicity of PVDF the membrane must be soaked entirely in methanol before equilibration in aqueous buffer Proteins might be transferring throu...

Page 27: ...27 Too low background Increase antibody concentration protein sample concentration Consult manual included with antibody detection kit Total protein detection Consult stain or detection kit manual...

Page 28: ...ormed by anyone other than CSL or an appointed distributor or representative are no longer under warranty from the time the unit was modified Units which have accessories or repaired parts not supplie...

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