Flow cytometry

Paul D. Boyer boyer at cs.uwp.edu
Thu Jul 27 14:17:31 EST 1995


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




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