Dan-
My flow cytometry experience at Caltech taught me that many mouse
monoclonals bind very well, though non-specifically to lymphocytes.
The best results I got was to block the non-specific binding with an
initial incubation in 1:10 normal mouse serum and inclusion of the same
in each of the antibody cocktails which follow. Good luck!
Paul D. Boyer
Assistant Professor of Biological Sciences
University of Wisconsin-Parkside
900 Wood Road, Box 2000
Kenosha, WI 53141-2000
ANDERSON at PRL.PULMONARY.UBC.CA wrote:
>> I am interested in performing flow analysis on murine splenocytes for
> T-cell antigens, B-cell antigens and enteroviruses. We have two
> antibodies, a mouse anti-virus monoclonal antibody and a rabbit
> anti-virus polyclonal. The CD antigens have worked extremely well
> under many conditions, however, we have had problems with
> non-specific staining with the anti-virus antibodies.
>> The mouse MAb is excellent for staining virus, however, in murine
> tissue the secondary gives the background. This occurs both with
> the conventional direct and indirect staining with respect to a labelled
> 2' anti-mouse antibody. The non-specificity of the rabbit antibody
> appears to be primarily in the rabbit antibody itself as there is no
> 2' Ab background staining.
>> If anyone has any comments or suggestions, I
> would enjoy discussing these issues.
>> Dan Anderson (danderson at prl.pulmonary.ubc.ca)
> The University of British Columbia
> The University of Nebraska Medical Center
