immunohistochemistry: L-selectin

[Rachel Teitelbaum]

I have done a dilution experiment, but used as the lowest dilution 1:20.  
I am using a secondary antibody, DAB is the readout.  I got staining of 
what I think are lymphocytes in spleen, but only in the red pulp, and 
don't know if this is "real" or not.  My negative control gave no 
staining.  I thought that the lung sample would have some positives, 
since its a mouse tissue that has had a respiratory infection.  These 
were paraffin sections, with an antigen retrieval protocol, that has 
worked for me in the past with other antibodies.  I can't use a FACS for 
this.  Any advice?

Should I stick with frozen sections?  Has anyone else had success with 
this antibody?

-Rachel

On Wed, 19 Feb 1997, Mark Haynes wrote:

> Rachel Teitelbaum wrote:
> > Hi Have you done a dilution experiment? Are you using a 2nd AB or a 
> biotin avidin system? What is the enzyme read out or are you using 
> fluorescence?  Can you stain lymphocytes and pick up positives on a 
> FACS?
> 
>