[Histonet] RE: Stupid rabbit antibodies

[Perry, Margaret]

Have you tried the Dako Rabbit envision?  We also tried the AbCam Expose Rabbit specific HRP and used Nova red from Vector  or the DAB in the kit depending on which antibody we used.  It is excellent , maybe even a little better than Dako's envision.  I did have to use blocks such as FC receptor block Innovex, Powerblock Biogenex and serum free protein block DAKO.  What I used depended on the antibody with some needing only the block provided in the kit.  Good Luck.

Margaret Perry HT(ASCP) 
Dept of Veterinary and  Biomedical services
Box 2175
South Dakota State University
Brookings SD 57007
605-688-5638


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Message: 17
Date: Tue, 22 Mar 2011 16:04:16 -0500
From: <sgoebel <@t> mirnarx.com>
Subject: [Histonet] Stupid Rabbit primaries!
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<D957F2A7D21959488C492A2680F9920A1C5FF5 <@t> svrexch.asuragen.us>
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So I haven't had to deal with rabbit polyclonal primaries in a long time
because I remember how much the background sucks with them.
Unfortunately the only available antibody is a rabbit polyclonal.  Does
anyone have any suggestions for how to eliminate the background?  I have
diluted almost to the point of the antigens not showing!  Thanks guys
and gals!!

 

Sarah Goebel, BA, HT(ASCP)

Histotechnologist

Mirna Therapeutics

2150 Woodward Street

Suite 100

Austin, Texas  78744

(512)901-0900 ext. 6912

 



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Message: 18
Date: Tue, 22 Mar 2011 16:13:04 -0500
From: "Sebree Linda A" <LSebree <@t> uwhealth.org>
Subject: RE: [Histonet] Stupid Rabbit primaries!
To: <sgoebel <@t> mirnarx.com>,	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<8C023B4AB999614BA4791BAEB26E2738010029DE <@t> UWHC-MAIL01.uwhis.hosp.wisc.edu>
	
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Do you know for sure that its the Ab and not endogenous biotin,
peroxidase or Phosphatase?  You can block for all those.

Otherwise there are a number of commercial blocking agents on the
market...try Biocare for a start (800)799-9499 or better yet, the
company that makes your antibody.

Good luck. 


Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
DB1-223 VAH
600 Highland Ave.
Madison, WI 53792
(608)265-6596