PeproTech
Human VEGF 165 Standard ABTS ELISA Development Kit
900-K10
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VEGF is one of the major growth factors that stimulate the formation of new (angiogenic) blood vessels. Measurement of VEGF production by tumor cells is important in the research focused on tumor angiogenesis and cancer metastasis. Since growth factor production by mammalian cells is relatively low (usually from picograms to nanograms per ml of conditioned medium or tumor extract), a sandwich (capture) ELISA format is required for the quantitative measurement of naturally expressed cytokines. We employed the PeproTech VEGF ELISA kit to measure and compare the levels of VEGF secreted by human tumor cells exhibiting different levels of metastatic dissemination in vivo.
Sandwich ELISA
Serum-free medium conditioned by tumor cells or primary tumor extracts. The ELISA kit from PeproTech contains all key components to measure VEGF produced by human tumor cells, namely the capturing antibody, the developing biotinylated antibody, recombinant VEGF to make a standard curve, bloking solution and the substrate to develop color reaction.
We closely followed the manufacturer's "ELISA: Sandwich ABTS Protocol", but with some modifications indicated below in the "tips" section.
The anti VEGF "capture" antibody could be coated at 0.5 microgram per ml, instead of recommended 1 microgram per ml. Coating of the "capture" antibody could be done at 4oC instead of RT. Washings could be done 3 times instead of recommended 4 times.
Yellow color substrate precipitates in the wells containing captured VEGF. Color development is measured with a spectrophotometer (ELISA plate reader) at 405 nm. The optical density is proportional to the levels of VEGF.
N/A
The image depicts the standard VEGF curve (the graph on the left) that is used to quantify VEGF concentrations in tumor cell samples (the graph on the right).
The kit is very specific, sensitive, and easily adaptable.
None detected.
Human VEGF kit "900-K10" from PeproTech is an excellent sandwich ELISA kit to quantify standard form of human VEGF (165).