----- Original Message -----

From: Cynthia Favara

To: 'Carrie Kyle-Byrne' ; Gayle Callis

Cc: 'HistoNet@Pathology.swmed.edu'

Sent: Tuesday, May 28, 2002 12:02 PM

Subject: RE: mouse monoclonal on mouse tissue

I agree it is definitely worth a try. I use a number of mouse Antibodies on mouse tissue. I think the key is what you are trying to detect and the tissue you are trying to detect the antigen in. A significant amount of background is caused by the anti Mouse IgG this can often be significantly decreased by a good perfusion. Many times a simple no primary control can give you the background noise and you can judge for yourself.

Cynthia Favara

-----Original Message-----
From: Carrie Kyle-Byrne [mailto:ckbyrne@exelixis.com]
Sent: Tuesday, May 28, 2002 11:15 AM
To: Gayle Callis
Cc: 'HistoNet@Pathology.swmed.edu'
Subject: Re: mouse monoclonal on mouse tissue

i beg to differ gayle.  i've done several mouse-on-mouse (most recent
notable is Ki-67 (clone MM1)) that have no or very little background using
our standard protocol (using Dako Envision+/DAB) with no additional
blocking.  i find it less time consuming if i try the aby first with no
special protocol and only go to the "mouse-on-mouse" stuff if absolutely
necessary.

Carrie Kyle-Byrne, BHS, HT(ASCP)
Assoc. Research Scientist II
Molecular Target Research

Exelixis, Inc.
170 Harbor Way
P.O. Box 511
South San Francisco
CA 94083-0511 USA

Phone: (1 650) 837-8023
Fax: (1 650) 837-7240
Email: ckbyrne@exelixis.com

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----- Original Message -----
From: "Gayle Callis" <uvsgc@montana.edu>
To: <histonet@pathology.swmed.edu>
Sent: Tuesday, May 28, 2002 9:12 AM
Subject: mouse monoclonal on mouse tissue

> The answer to the question is yes, background will be a huge problem.
>
> Try a mouse on mouse kit, Zymed, Innogenex, Scytek, Vector all have
special
> kits containing special blockers. I hope I haven't missed a vendor, if so,
> speak up if you have the kit.
>
> Another alternative is DAKO ARK kit, based on biotinylation of the primary
> antibody and their special blocking, etc.
>
> Some people have tried more than one kit for success, they do vary - but
> you may be lucky with first one you try.
>
>
>
>
> Gayle Callis
> MT,HT,HTL(ASCP)
> Research Histopathology Supervisor
> Veterinary Molecular Biology - Marsh Lab
> Montana State University - Bozeman
> 19th and Lincoln St
> Bozeman MT 59717-3610
>
> 406 994-6367 (voice mail)
> 406 994-4303 (FAX)
>
> email: UVSGC@montana.edu
>
>