FACS of stimulated human PBMC - (Jul/11/2006 )
Who has the experience in FACSing of stimulated human PBMCs?
Our problem is as follow:
the staining on unstimulated cells looks great, but after the stimulation shifted mIgG1-FITC to the right till 10²-10³. So we have no or only little staining with our target...
we block with 10% FCS or AB-serum....
-Katja W.-
Maybe there is an upregulation of Fc receptors? I could see that happening is your "stimulation" was immunological in nature.
Instead of just blocking with FCS maybe you could try using some 2.4G2 antibody. It binds Fc receptors and blocks them. Just pretreat with it beofe all antibody stainings.
maybe that will help.
-Mountainman-
QUOTE (Mountainman @ Jul 12 2006, 03:44 AM)
Maybe there is an upregulation of Fc receptors? I could see that happening is your "stimulation" was immunological in nature.
Instead of just blocking with FCS maybe you could try using some 2.4G2 antibody. It binds Fc receptors and blocks them. Just pretreat with it beofe all antibody stainings.
maybe that will help.
Instead of just blocking with FCS maybe you could try using some 2.4G2 antibody. It binds Fc receptors and blocks them. Just pretreat with it beofe all antibody stainings.
maybe that will help.
Thank you for your answer!
Do you know any anti-human CD16/CD32 antibodies? Because 2,4G2 seem to be anti-mouse...
-Katja W.-