Jos, When speaking of quantitative analysis of stem cells, I would not substitute CD34-PE with any other fluorochrome. We tried to use CD34-APC in our ISHAGE-like procedures ( I am not even saying PerCP) and saw a difference right away. If you plan just to characterize the population of interest, we use CD34-PerCP-Cy5.5: we like it just fine and we have data. And PE-Cy7 may be a good choice, though I have no data to support this statement. Irina Irina Grigorieva, PhD Director, Flow Cytometry Laboratory Northside Hospital, Atlanta, GA (404)- 851-6541 e-mail: irina.grigorieva@northside.com ________________________________ From: Jos Bosch [mailto:j.a.bosch@bham.ac.uk] Sent: Thursday, December 20, 2007 10:11 AM To: cyto-inbox Subject: CD34 CD133 Dear Flowers, Am setting up a panel on a 6-color FACScanto to characterize stem cells in peripheral blood, and running into the usual planning problem on what fluorochromes to assign to particular antibodies. The main hassle is caused by the fact that CD133 (miltenyi) and KDR (R&D) are only available in PE and APC, so have to work around these. To make a long story short, there are two options I seem to be left with: 1) Substitute CD34 PE (clone 8G12, BD) for PerCP. Arseniev (1999) concluded PerCP was not really bad, but not really great either. What is your experience?? Was unable find any systematic comparison in the literature that looked at PE-Cy7 or APC-Cy7. Does anyone have good validation data? 2) Alternatively, I can buy CD133 biotin and label it with FITC. Does anyone have good experience with that approach? What can I expect with regard to signal strength, background, and prep hassle? Your advise is very much appreciated! Kind wishes, Jos Jos A. Bosch, PhD Lecturer in Exercise and Behavioural Immunology School of Sport and Exercise Sciences University of Birmingham Birmingham B15 2TT United Kingdom T: (+44) 0121 4148105 F: (+44) 0121 4144121 http://www.sportex.bham.ac.uk/staff/boschja CONFIDENTIALITY NOTICE: This electronic mail transmission has been sent by Northside Hospital. It may contain information that is confidential, privileged, proprietary, or otherwise legally exempt from disclosure. If you are not the intended recipient, you are hereby notified that you are not authorized to read, print, retain, copy or disseminate this message, any part of it, or any attachments. If you have received this message in error, please delete this message and any attachments from your system without reading the content and notify the sender immediately of the inadvertent transmission. There is no intent on the part of the sender to waive any privilege.Received on Sat Dec 22 15:18:00 2007
This archive was generated by hypermail 2.1.8 : Wed Jan 31 2007 - 03:12:00 EST
