[Cytometry] Anti-CD3 activation for phosflow

Kit Fuhrman kit.fuhrman at pathology.ufl.edu
Fri Apr 5 18:06:06 EDT 2013


The treg inspector beads from miltenyi might work a little better for that application.    They're supposed to fall off real easy with a little bit of pipetting then easily gated out by SSC.  The dynabeads can show up in some of the other channels.  I've had them show up as double negatives and make the data a little messy.




On Apr 5, 2013, at 5:40 PM, "Prussin, Calman (NIH/NIAID) [E]" <CPRUSSIN at niaid.nih.gov<mailto:CPRUSSIN at niaid.nih.gov>>
 wrote:

Flowees!

Has anyone experience with or know of a reference for using Dynabeads® Human T-Activator CD3/CD28 to activate T cells or PBMC for phospho-flow? The phosphorylation events we are looking at (mTOR downstream effectors) take a few hours to occur.

Other then the beads changing cell scatter is there a problem with using them in an LSRII?

http://products.invitrogen.com/ivgn/product/11161D

Thanks,

Calman
______________________
Calman Prussin
Laboratory of Allergic Diseases
National Institute of Allergy and Infectious Diseases/ NIH
www.tinyurl.com/prussin1<http://www.tinyurl.com/prussin1>

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