Regarding primary antibody binding in western blot

Jayakumar, R via methods%40net.bio.net (by R.Jayakumar from roswellpark.org)
Wed May 16 15:08:08 EST 2012


I have also observed people probing multiple PVDF blots separated by a nylon mesh to prevent the blots from sticking to each other, in a box that is just larger than the blot itself.  I thought that was a neat idea.  You can try that way too. 
Jay



-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu [mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of Irit Rappley
Sent: Wednesday, May 16, 2012 1:34 PM
To: divya teja
Cc: methods from magpie.bio.indiana.edu
Subject: Re: Regarding primary antibody binding in western blot

Hi Divya,

Yes, you can certainly put 2 membranes in the same box with your primary antibody. Just make sure that the membranes are back to back, so that the protein side of each membrane is facing outward to the antibody solution.

Good luck!
Irit


On May 16, 2012, at 6:03 AM, divya teja wrote:

> hello,
> 
> Can I use 2 PVDF  membranes at the same time in single box containing 
> primary antibody in 10ml 1x PBST while doing western blot?
> 
> Will the primary antibody bind properly or gets probed properly, if I 
> use 2 membranes same time in the single box????????
> 
> waiting for the reply
> 
> 
> regards
> divya
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> Methods from net.bio.net
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