I need know which is the best (with references if possible) control label to use for FACS as control group for cells obtained by enzyme digestion process of porcine vein tissue. Is any way to create a control label constitutionally express in all cells that allows me to create a control to compare with the interested labelled cells?
VEGF receptors (1,2,3) perhaps, but remember to choose antibody for porcine sections.
It is important to understain from which type of cells you serch to identify: if epithelial you can use a vimentin antibody.
BD Bioscience offers reliable antibodies for FACS:VEGF might be a candidate.
But you'll have to cintact customer service to get information with respect to species cross reactivity
http://www.bdbiosciences.com/nvCategory.jsp?action=SELECT&form=formTree_catBean&item=746070
On the other hand abcam also offer reliable antibodies : eg. VEGF
http://www.abcam.com/vegf-antibody-ep1176y-ab52917.html
Also check for cross reactivity.
On the other hand, another supplier might offer VEGF primary antibodies which you might label as you wish.
Thank you both for the useful information, i will look into that. Seems the VEGF is the target. Thanks again.
if your question is how do I know that I got a population enriched with endothelial cells, the answer is to use an Ab against endoglin (CD105) or MECA . those are expressed only on ECs... Flk Flt VEGF are expressed in many many other cells...
If you are determining any protein changes in the endothelial cells, you can tag for eNOS, since its an internal protein. As metioned above by others, you can also tag for VEGF receptor, but since you are trypsinizing, you may be questioned. It depends on what you are measuring?
Good luck
Regards
Paul
If you search a cytoplasmatic protein remember to permeabilize membrane before.
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