HELP !!---Endocytosis assay

BDam815174 bdam815174 at aol.com
Thu Sep 23 22:11:33 EST 1999


Hi All!

I am trying to perform an endocytosis assay...
After I bind my labeled antibody to my receptor protein (4C) and wash
off the excess antibody, I put the cells at 37C for different
time points.  I then take an aliquot of cells and try to remove any
anitbody still bound to the cell surface by washing my cells
in PBS (pH2).  This should give me the amount of internalized antibody.
However everytime I was the cells with the acid buffer I lyse them...

What am I doing wrong?
Every protocol I read on this assay says to wash in acid
buffer to remove any external bound antibody but this is killing
my cells !!!!!

HELP!!!!

Please reply to:
bdamania at world.std.com

Thanks a lot !!!




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