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HUVEC surface antigen staining


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#1 gdhk

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Posted 25 March 2009 - 06:38 PM

Hi,

Can anyone help stain HUVEC cell surface antigens. I have used Trypsin & a non-enzymatic cell dissociation buffer for detachment from the collagen monolayer. I have been washing the cells with PBS/0.1% (PBS pre-detachment) and using anti-vegfr2, anti-CD40 and anti-vcam antibodies for staining (incubation on ice). I have not been able to get a positive fluorescence for any of the antibodies. Does anyone have experience staining HUVEC surface antigens? Any help would be greatly appreciated.

Thanks.
GDHK

#2 CKtong

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Posted 04 April 2009 - 12:19 AM

Hi,

Can anyone help stain HUVEC cell surface antigens. I have used Trypsin & a non-enzymatic cell dissociation buffer for detachment from the collagen monolayer. I have been washing the cells with PBS/0.1% (PBS pre-detachment) and using anti-vegfr2, anti-CD40 and anti-vcam antibodies for staining (incubation on ice). I have not been able to get a positive fluorescence for any of the antibodies. Does anyone have experience staining HUVEC surface antigens? Any help would be greatly appreciated.

Thanks.
GDHK


Hi,
I had working various cell for flow(include HUVEC). so far not expericences any problem.
I suggest u to use non- encymatic method. beside, what antibodies u used? (which company? flourecent labelled? Or need any secondary stain?)




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