Antibody cocktail for Western blots?

John Thompson jrt at home.com
Sat Oct 10 21:42:40 EST 1998


In priciple there's no reason you can't mix antibodies to light up
multiple bands.   But if they're not monoclonals, the additive effect
of all the preimmune bands could obscure what you want to see.

Regards,
John Thompson


Arul Jayaraman <jayarama at helix.mgh.harvard.edu> wrote:

>Hi
>I am studying the acute phase response in post burn situations using two
>dimensional gel electrophoresis. I am interested in a set of 6 acute phase
>proteins.  While I can do Western blots to identify each protein
>individually on 2D gels, this means that I need to run six 2D gels for
>each sample. What I really need is to be able to detect as many proteins
>as possible on the same gel. My questions are:
>1. How many times can one strip and reprobe nitrocellulose membranes for
>Westerns?
>2. Is it possible to have 'antibody cocktails' (antibodies for many
>proteins in one mix) and use that for probing? All the primary antibodies
>that I have are from goat and I use HRP detection with the seconday
>conjugated antibody
>
>Thank you
>
>Arul Jayaraman
>jayarama at helix.mgh.harvard.edu
>-----------------------------------------------------
>Arul Jayaraman
>Center for Engineering in Medicine
>Massachussetts General Hospital
>
>Mail to:
>Shriners Burns Research Center
>One Kendall Square, Building 1400 West
>Cambridge, MA 02139
>
>(617) 374-5625 / (617) 374-5611 (Phone)
>(617) 374-5665 (Fax)




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