You need to either pre-soak or wash the blood before adding the antibody or it is aborbed by elements in the plasma. We wash the blood three times with PBS + 1%FBS and then stain and they come out great. You should be able to test it on normal whole blood to make sure it is working right and you should get a 50/50 split on CD19 cells. Brian Newsom Director, Flow Cytometry Center for Cell and Gene Therapy Baylor College of Medicine -----Original Message----- From: Sebastian, J, Gelderbloem, Mr [mailto:sjg@samiot.uct.ac.za] Sent: Wednesday, July 14, 1999 10:10 AM To: cyto-inbox Subject: Re: CLL PHENOTYPE Dear Flowers I have a slight problem and I am not sure what the solution is. We use polyclonal Kappa and Lambda from BD for our chronic leukaemia panel. We have been setting the samples up using whole blood and for some or other reason the Light Chains are coming up negative everytime. We only bitmap lymphoid cells. Is there anyone with some advice. Thanks Sebastian
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