IP Problem

Jayakumar, R via methods%40net.bio.net (by R.Jayakumar from roswellpark.org)
Fri Jul 11 16:41:45 EST 2008


Not very difficult at all.  Use a different antibody (for e.g. Mouse)
for IPing the protein down and use another one for western detection
(for e.g. Rabbit).  Then the heavy chain of mouse will not be visible
when we use anti-Rabbit secondary for western detection.

Jay

-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu
[mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of Dr Engelbert
Buxbaum
Sent: Friday, July 11, 2008 2:37 PM
To: methods from magpie.bio.indiana.edu
Subject: Re: IP Problem

Am 10.07.2008, 13:43 Uhr, schrieb John Leonard <jleonard.iii from gmail.com>:

> Hello all,
>
> I'm looking to perform an IP Western of a protein that runs at around 
> 50-55kD on an SDS-PAGE gel - approximately the same as my antibody's 
> heavy chain.  As such, it could be very difficult to detect presence 
> of my protein


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