From: colette charland (ccharlan@moose.uvm.edu)
Date: Tue Dec 19 1995 - 09:34:19 EST
As promised, here is my second request....any takers?
Thanks colette
>Date: Mon, 11 Dec 1995 17:16:01 -0500
>X-Sender: ccharlan@moose.uvm.edu
>To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
>From: ccharlan@moose.uvm.edu (colette charland)
>Subject: Platelet analysis/Va antibody
>
>Hello out there!!
> I have an investigator who is labelling a Va antibody with Cy5 and
would
>now like to use it to tag activated platelets. My understanding is that the
>Va is located within granules, which upon activation move to the surface go
>through exocytosis and release the Va. She tells me that the Va "jumps
>onto" (this is a direct quote) and attaches to Xa on the surface of the
>platelets. Is this correct? Are there any Ca++ requirements for this
>reaction.
> I know that the platelets are in fact being activated because we
>have been using cd62 as a positive control. Unfortunately, she is on a
>learning curve as far as the tagging procedure with the Cy5, and I am on a
>learning curve in terms of using the dual laser set-up on my Elite. I do
>however beleive that I am where I need to be perhaps with a little
>tweeking. Any suggestions or pertinent info would be greatly appreciated.
> If this doesn't work, we are thinking of biotinylating the reagent
>and then using SA-670 (or whatever). Is this possible on will the molecule
>be too large for the platelet?
> Any advice from someone with knowledge of the requirements or
>physiology of the activation pathway would be greatly appreciated
>
>
> THANKS COLETTE CHARLAND
> ccharlan@moose.uvm.edu
>
>
>
>
>
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