Sharon,
 
I know a group that uses confocal but do not know it very well myself.  Can you give me a more detailed explanation of why confocal would be useful ... and suggestion for cell surface markers that can be used?
 
Thank you very much.

Sharon Cooperman  wrote:
You could do confocal microscopy - that would unequivocally 
demonstrate thqt the antigen is on the cell surface, particularly if 
you co-stain with a cell surface marker.

Sharon

>Hello,
>
>How would one go about determining if your specific antigen (say for 
>IHC) is located on the cell surface? Right now it seems that the 
>antigen is staining everywhere. (I am running a sheep anti-mouse 
>'antigen' and using a rabbit anti-sheep biotin conjugated with ABC + 
>DAB for detection. (My negative is normal sheep IgG -- is 
>negative). Is there a way through immuno or other that I can 
>specifically say that the antigen is present on the cell surface? 
>Are there general cell surface markers that I can use along with my 
>primary? Thank you all.
>
>
>
>
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-- 
Sharon Cooperman 
NIH, NICHD, CBMB 301.435-8417
Building 18T, room 101 301.402-0078 fax
Bethesda, MD 20892

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