Hi, I would like to know if any of you have a protocol to isolate proteins from acrylamide gels. I want to extract proteins and then run them again for Western blot
Regards,
Monsa
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#2
scientist
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Posted 27 January 2005 - 10:02 AM
What are you doing to the protein the first time round?
Coomassie staining or something else?
Coomassie staining or something else?
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Mercifully free of the ravages of intelligence - Time Bandits
Mercifully free of the ravages of intelligence - Time Bandits
#3
snolan6
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Posted 02 February 2005 - 01:03 AM
You can use coomassie to visualize in order to cut out your band, but avoid using methanol and acetic acid during the stain or destain procedure - this will fix the protein into the gel and reduce the yield during protein elution. Use water as the solvent instead...
I think you have 2 options for elution
1 - electroelution
example : http://www.fermentas...telutatubet.htm
2 - ultrafiltration
example: http://www.pall.com/34696_2273.asp
someone might be able to give a "from scratch" protocol though. Im a fan of kits...
good luck
I think you have 2 options for elution
1 - electroelution
example : http://www.fermentas...telutatubet.htm
2 - ultrafiltration
example: http://www.pall.com/34696_2273.asp
someone might be able to give a "from scratch" protocol though. Im a fan of kits...
good luck
