Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

TCA Precipitation to remove albumin

TCA Precipitation albumin

  • Please log in to reply
6 replies to this topic

#1 Anupa

Anupa

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 18 August 2012 - 09:12 AM

I am doing a project with human serum, as i need to remove albumin., i tried TCA preciptation, according to a new protocol saying that TCA preciptation can reduce albumin content.. i used 4 volumes of TCA/acteone. after the entrie procedure i got a pellet. the problem is i tried to dissolve the pellet in 1M tris pH 6.8 but I cudnot dissiolve. After TCA perciptation i need to go for protein estimation by lowrys method then SDS-PAGE then 2D gel then mass spectrometry.. I have read tht we can dissolve pellet in 2X SDS-Loading buffer... But as i need to go for protein estimation by lowry's i don't think so i can go for SDS loading buffer... Pls do just me a method were i can continue through my entire project....

Anu

#2 Curtis

Curtis

    Metaller Scientist

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,112 posts
58
Excellent

Posted 18 August 2012 - 08:05 PM

I would dissolve in RIPA buffer. It never affected my Bradford assay reading, even after addition of protease inhibitor.

#3 BioMiha

BioMiha

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 229 posts
9
Neutral

Posted 19 August 2012 - 11:51 PM

It it were me, I would not use TCA to remove albumin, because it non-selectively precipitates all proteins. There are kits available for albumin depletion, which are used prior to 2D electrophoresis and I have heard they're quite OK.
As for dissolution of your pellet, I would go with SDS buffer and BCA or Bradford for quantification.

#4 Anupa

Anupa

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 20 August 2012 - 07:39 AM

thanks for the suggestion but i have read an article that said it would reduce the content of the albumin present in the sample please do help me in this d suggesting a way out... like an affordable albumin removal kit.

Edited by Anupa, 20 August 2012 - 07:39 AM.


#5 BioMiha

BioMiha

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 229 posts
9
Neutral

Posted 21 August 2012 - 02:27 AM

There are a number of products on the market. Why don't you just do a Google search for a kit and try to find something that's within your budget. I know of kits from Qiagen and GE, but I don't know if they are within your price range.

#6 Anupa

Anupa

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 21 August 2012 - 08:04 AM

I know about those Kits.... And most of them is beyond my budget.. That is when I found about this TCA precipitation...

#7 cbf88

cbf88

    member

  • Active Members
  • Pip
  • 14 posts
0
Neutral

Posted 29 September 2012 - 01:27 AM

dissolve in urea lysis buffer and bca assay it





Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.