ECM Autofluorescence blues - any solutions?

From:Gail Donegan

Hi everybody,

Thanks to everyone for the help and suggestions regarding what I mistakenly 
assumed was non-specific binding of my secondary antibody. Now I find I 
have a different problem altogether...

The extracellular matrix (ECM) laid down by my human smooth muscle cells 
fluoresces red at 594nm (so fluorescently staining with an antibody that 
fluoresces red is pointless) and which also fluoresces green at 495nm (so 
using a green fluorescing antibody is no good either).

I've tried using 100mM - 500mM Glycine, which worked for Andrea who kindly 
responded to my plea last time I was in touch.

Does this mean that fluorescently staining adsorbed ECM is not possible or 
is someone out there doing it?

Any suggestions gratefully received,

Thanks,

Gail



Gail Donegan
Department of Clinical Engineering,
The University of Liverpool,
Duncan Building,
Daulby Street,
Liverpool,
L69 3GA.
U.K.

Tel:    +44(0)151 706 5198
Fax:    +44(0)151 706 5803




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