Fc Blocking - Mutz-3

Joost Schuitemaker j.schuitemaker at iqproducts.nl
Wed Sep 19 14:23:46 EDT 2007


Hi Cody,
 
I used to work with the line. Best thing to block with is human serum. The FcR has highest affinity for those antibodies. Add this to your incubation medium too. Second choice would be a mixture of Fc blocking antibodies, like Rachel suggests. Last option is to block with mouse Ig or mouse serum. But this only works when you don't have second step incubations in you staining.
 
Regards,
 
Joost
 

-----Original Message-----
From: Cody Allison [mailto: <mailto:Cody.Allison at med.monash.edu.au> ]
Sent: Mon 9/17/2007 5:05 PM
To: cyto-inbox

Hi,

I am currently working with the DC-like precursor cell line, Mutz-3. I am having some
trouble with non-specific binding of my isotype control antibodies. I am using FITC
(IgG2a), PE (IgG1) and PECY5 (IgG1) - conjugated antibodies.

I do block the cells with FCS, but was wondering whether anyone has encountered this
problem with, or has any specific blocking protocols when using Mutz-3. Any advice would
be greatly appreciated.

cheers
cody



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