VEGF for western blot - (Mar/13/2009 )

Hello,

I was wondering whether anyone has done western blots to look for VEGF in human cells? If you have, then did you collect the media from the cells, precipitate the proteins using 80% acetone and then ran a sample of that or did you collect the lysates? I have tried it both ways, although I am not surprised that I didn't see VEGF in whole cell lysates as it is being secreted from the cells. Any help is appreciated.

Thanks.

A colleague in my lab used ELISA to sucessfully detect it

yobou on Mar 13 2009, 11:54 PM said:

I'm trying to detect VEGF in media from HUVECs using Quantikine ELISA from R&D systems but have had no success. Very frustrating, and Western blots of HUVEC lysate using Santa Cruz Ab also doesn't detect anything.

Has anyone had any luck with VEGF in HUVECs?