1 reply to this topic

[gdhk]

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

[CKtong]

gdhk, on Mar 26 2009, 10:38 AM, said:

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?)