Re: [Histonet] mouse bile duct epithelium

From:Gayle Callis

How about trying another mouse to mouse kit that does not involve
biotinyated secondaries.  

The key to mouse to mouse blocking in Scytek's kits are the two blocking
reagents, one could use their blockers which they will separately from a
kit, use your mouse to mouse primary, and plug in a secondary that is
directly conjugated to HRP, eliminating biotin altogether. That could be
another possibility. Basically you are setting up your own mouse to mouse
kit with some superb blocking reagents. 

 DAKO's avidin biotin block, I have heard, is a higher concentration to
deal with CSA or tyramide amplification involving biotin, and may work
better unless you are already using DAKO's and increase the time of the
avidin/biotin blocking to insure endog biotin in liver is zip. 

Good luck  

At 01:24 PM 1/8/2004 -0700, you wrote:
>here is my story:
>i am trying to label ms bile duct epithelium with mouse anti-mouse
>cytokeratin 7 and 19 (sounds odd, but that is what the investigator tells me
>it is, not even rat anti-mouse) they tried the anti-human ck7&19 and it did
>not cross react with the mouse.  I have the reagents to biotinylate these
>msbiotin is a huge problem even with the most aggressive A/B blocking.  Is
>there another possible rabbit antibody I could use that would label ms bile
>duct epithelial cells?  Would factor VIII, cd31, or cd34 stain these cells?
>I know that the previous mentioned markers are for endothelial cells but I
>am just hoping they might work?????  I have not been able to find any other
>say rabbit anti-ms ck7 or ck19 out there, has anyone else.  How about the
>new rabbit monoclonals, is anyone making ck7/ck19?  By the way I have frozen
>and ffpe bile duct samples, so I can use either.
>Please advise.
>Patsy Ruegg
>
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)



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