Weiwen, It sounds like your friend is using an anti-CD59 and then using a complete or F(ab)2 secondary fluorochrome conjugated antibody for detection. When working with red cells it is important to use a Fab secondary antibody otherwise you will get agglutination (as you have described). We have done a lot of flow work on RBCs and have found the secondary antibodies from Jackson ImmunoResearch very good. Or you could use a directly conjugated anti-CD59. Silenus has a very good one http://www.silenus.com.au/. Jenny Bryant Flow Cytometry Australian Red Cross Blood Service - NSW/ACT 153 Clarence St Sydney, NSW 2000, AUSTRALIA .._|\ Ph: 61 2 9229 4341 / \ FAX: 61 2 9229 4521 \_.-._/<<<<<< mailto:jbryant@arcbs.redcross.org.au -----Original Message----- From: Weiwen Shen [mailto:shenww@public3.sta.net.cn] Sent: Friday, 17 November 2000 0:42 To: cyto-inbox Subject: about CD59 detection Cheers flowers, I have a friend trying to detect CD59 expression on erythrocytes in PNH patients and healthy people. He found that the erythrocytes were aggregated, and he got two peaks in healthy controls. We wonder whether there is some mistake in sample treatment. So we expected the experts here could help us. Weiwen Shen
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