RE: Annexin binding on platelets

Jim this is essentially the method I use for platelets (I have a few extra bells and
whistles). For an "isotype" add 5ul of 50mM EDTA and 25ul HEPES buffered saline instead
of the 30ul for the labelling solution below. Bones of the method are published in
Transfusion:- Garwood et al Transfusion. 2003 Sep;43(9):1238-47, but should be published
for platelets soon.

best wishes Craig

Labelling solution
5ul 10x Hanks buffered salt solution
10ul FITC:annexin V (Caltag Medsystems)
30ul HEPES buffered saline
5ul of platelets at 100 x 10^9/L (dilute in HEPES buffered saline)

Method
Incubate for 20 min Add 450ul Hanks buffered salt solution(stable for another 60 min)
Analyse 





Craig Turner PhD
Senior Component Development Scientist
National Blood Service
Tel 01277 306194
Fax 01277 306043

-----Original Message-----
From: MIME :KurtzJ@usa.redcross.org Sent: 15 July 2004 20:52
To: cyto-inbox
Subject: Annexin binding on platelets


I was wondering if someone had a procedure to measure annexin V on platelets.  I need to
measure annexin binding using
platelet concentrates.	Thanks,  Jim Kurtz




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