Quantification of VEGF peptide levels in human breast cancer cells. HELP! - (Dec/16/2008 )

Hi ,
I am currently doing a study on anti- angiogenic study of several natural product extratcs. One of the highlights of this study is also to see how these extracts effect the Vascular Endothelial Growth Factor (VEGF) inhibition in human breast cancer cells, particularly MDA-MB-231 cell lines. therefore i would like to quantify the levels of VEGF peptide levels produced by these cells. Can anyone tell me if this is possible and if so, how should i approach this? k have read several articles usinf a VEGF-ELISA assay however explanation on the methodology is very vague. Do cells grown in a flask secrete VEGF ? or do i need to induce the secretion of VEGF in this flask grown cells? i would be grateful if someone cld help me.
thanks in advance.

I found this kit:

http://www.raybiotech.com/manual/ELISA/ELM-VEGF-001.pdf


Perhaps, they can help you with inducing secretion information

Dear pronana,

if you want to measure the amount of VEGF being secreted by your cells, you need to grow the cells in a flask (about 70-80% confluent), then collect the media and perform ELISA or EIA using the media. RIA can also be done, but that would need radioactivity etc etc,... I would stick on to just ELISA.. I hope this has answered ur query.

Good Luck
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Dear molecular_medicine,
How do i perform the ELISA what do i use to give the colored product for in order to measure it spectrometrically? antibodies? do you have the protocol for this assay? If you dont mind me asking, what is EIA?

Thanks in advance for your time

http://www.rndsystems.com/pdf/qve00b.pdf check this out!!!!!!!!!!!! good luck....