HUVEC surface antigen staining - (Mar/25/2009 )
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
-gdhk-
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
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?)
-CKtong-