[Protein-analysis] Protocol for human CD38 WB

Morales, Victor via proteins%40net.bio.net (by Victor_Morales from URMC.Rochester.edu)
Sat Feb 2 16:30:54 EST 2008


 Connie:
Have your mab's been tested for western blotting? Some antibodies only
recognize the native protein conformation and cannot bind to the
denatured forms in the western blot.
Victor

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Today's Topics:

   1. Requesting protocol for human CD38 western	analysis (Connie
Lam)


----------------------------------------------------------------------

Message: 1
Date: Fri, 1 Feb 2008 12:43:19 +0800
From: "Connie Lam" <boconnie from gmail.com>
Subject: [Protein-analysis] Requesting protocol for human CD38 western
	analysis
To: proteins from magpie.bio.indiana.edu
Message-ID:
	<93f327860801312043q76210882xba9eb6ad9c5b0512 from mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

 Dear all,

*Requesting protocol for human CD38 western analysis*

I are studying human CD38 with different hCD38 monoclonal antibody
T16/IB4.
I can use the Ab to stain hCD38-transfected HEK293 cells. When I used
T16/IB4 for western, no bands were observed.

 Here is my protocol. I am using the RIPA buffer + protease inhibitor to
lyse the cells and run on 10% SDS-PAGE gel. After transfered onto
nitrocellulose membrane and blocked with 5% skim milk, the membrane was
incubated with one of the Ab (4ug/ml) overnight at 4oC followed by
anti-mouse-HRP for 1 h at room temperature. I am looking for other
protocols for sample preparation and western for hCD38?

Thanks.

Connie


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