Dear all, I am trying to label human peripheral blood lymphocytes to detect expression of the common gamma chain of the IL-2, IL-4..etc receptor. I normally just lyse out red cells from whole blood and wash white cells with PDS / Azide (0.1%)/ BSA (0.1%) before labelling. I'm using a rat-anti human monolconal-Biotin and then Streptavidin-Fitc, and am having problems with consistent labelling. That is, sometimes the lymphocytes label, othertimes not. I've tried various temperatures (RT, 4C, on ice), and tried titering the antibody over a wide range to optimise labelling, but haven't been able to come up with a consistently good result. Does anyone have any experience with trying to label this antigen? Or is it more likely to be a problem with the antibody? Many thanks Belinda Kramer Oncology Research Unit The Children's Hospital at Westmead Australia Belindak@chw.edu.au ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of the Childrens Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. **********************************************************************
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