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Protein isolation from acrylamide gels


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#1 monsa

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Posted 26 January 2005 - 02:53 PM

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

#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?
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#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




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