If the anti-Mouse IgM is a polyclonal Ab it may have some cross-reactivity to human IgM. You could do your staining in the presence of human serum. (there is no perfect Fc block for human, this might also be your best Fc block) In addition you could use a monoclonal anti-mouse kappa as the secondary reagent. They tend to show little staining of human cells. Alan Stall BD Biosciences ======================================================== "Elkabetz, Yechiel/Sloan-Kettering Institute" <elkabety@mskcc.org> 10/28/05 01:32 PM To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> cc: (bcc: Alan Stall/SDCA/BDX) Subject: Reducing background when staining with 1st and 2nd antibodies Hi, I am staining human cells with a mouse IgM as 1st antibody and then FITC-anti-mouse IgM as a 2nd. What can I do to reduce background? This is because I get a non specific staining of the secondary, which makes the negative population shifted to be very close to the positive. Maybe an FC block? Thanks Techie ----------------------------------------- ************************************************** IMPORTANT MESSAGE FOR RECIPIENTS IN THE U.S.A: This message may constitute an advertisement of BD group's products or services or a solicitation of interest in them. If this is such a message and you would like to opt out of receiving future advertisements or solicitations from this BD group, please forward this e-mail to the optoutbygroup@bd.com. ************************************************** This message (which includes any attachments) is intended only for the designated recipient(s). It may contain confidential or proprietary information and may be subject to attorney-client privilege or other confidentiality protections. If you are not a designated recipient, you may not review, use, copy or distribute this message. If you receive this is in error, please notify the sender by reply e-mail and delete this message. Thank You ************************************************** Corporate Headquarters Mailing Address: BD (Becton, Dickinson and Company) 1 Becton Drive Franklin Lakes, NJ 07417 U.S.A.Received on Tue Nov 1 16:38:00 2005
This archive was generated by hypermail 2.1.8 : Sat Jan 14 2006 - 22:03:57 EST
