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
- - - - -

BSA concentration for blocking


  • Please log in to reply
5 replies to this topic

#1 science noob

science noob

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 281 posts
20
Excellent

Posted 14 December 2011 - 04:16 AM

How much BSA is enough to0 sufficiently block a membrane?

1%? 5%? 10%?

#2 GNANA

GNANA

    GNANA

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 209 posts
9
Neutral

Posted 14 December 2011 - 06:58 AM

5% is enough .....
I would prefer being perfectionist rather than a passionist in Research.

I always had an alternate hypothesis....

#3 lab rat

lab rat

    Why does a science forum not have pictures of mice and rats?

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 245 posts
7
Neutral

Posted 14 December 2011 - 10:12 PM

You can use 1%, if you block longer with shaking. I once made a mistake making TBS + BSA (0.5%) and didn't even notice until the liter was half gone. The background wasn't significantly different, probably because I blocked overnight at 4C.
42..."An immutable fixed-precision number of unlimited magnitude." <a href="http://en.wikipedia....amming_language)" target="_blank">http://en.wikipedia....amming_language)</a>, accessed 25June2009.

#4 BioMiha

BioMiha

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 236 posts
9
Neutral

Posted 14 December 2011 - 10:57 PM

I myself have generally had bad results with BSA blocking (high background). We use milk instead.

#5 science noob

science noob

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 281 posts
20
Excellent

Posted 15 December 2011 - 03:44 AM

I myself have generally had bad results with BSA blocking (high background). We use milk instead.


I found the opposite when doing a particular antibody. BSA blot was sparkling clean whilst milk gave alot of grey spots, smears and background after adding ECL. I think it has something to do with the primary antibody. I was detecting a phospho-protein. Milk was fine with the native form.

BSA is quite expensive so I was wondering if 1% does the job, it'll be good to save up on BSA stocks.

#6 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,826 posts
413
Excellent

Posted 15 December 2011 - 12:05 PM

Milk is full of phospho-proteins, so it tends to give dirty blots when used with an anti-phospho-protein antibody.




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

©1999-2013 Protocol Online, All rights reserved.