Platelet analysis/Va antibody
colette charland (ccharlan@moose.uvm.edu)
Mon, 11 Dec 1995 17:16:01 -0500
Hello out there!!
I have an investigator who is label 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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