Abcam, ab273164, Human VEGF-165 ELISA Kit

Size: 1 x 96Tests
Human VEGF-165 ELISA Kit is a Sandwich (quantitative) ELISA for the measurement of Human VEGF-165 in Human in Cell Culture Media, Biofluids samples.
Key facts
Detection method:Colorimetric,
Sample types:Cell culture supernatant, Plasma, Serum,
Reacts with:Human,
Assay type:Sandwich (quantitative),
Sensitivity:= 15.9 pg/mL,
Range:31.25 - 1000 pg/mL,
Assay time:3h 45m,
Assay Platform:Microplate (12 x 8 well strips)

Product details:
Human VEGF
ELISA kit (ab273164) is for the in-vitro qualitative and quantitative determination of VEGF
in supernatants, buffered solutions or serum and plasma samples. This assay will recognise both natural and recombinant human VEGF
A capture antibody highly specific for VEGF-A has been coated to the wells of the microtiter strip plate provided during manufacture. Binding of VEGF165 in samples and known standards to the capture antibodies is completed and then any excess unbound analyte is removed.
During the next incubation period the binding of the biotinylated anti-VEGF165 secondary antibody to the analyte occurs. Any excess unbound secondary antibody is then removed.
The HRP conjugate solution is then added to every well including the zero wells, following incubation excess conjugate is removed by careful washing.
A chromogen substrate is added to the wells resulting in the progressive development of a blue coloured complex with the conjugate. The colour development is then stopped by the addition of acid turning the resultant final product yellow. The intensity of the produced coloured complex is directly proportional to the concentration of VEGF165 present in the samples and standards.
The absorbance of the colour complex is measured and the generated OD values for each standard are plotted against expected concentration forming a standard curve. This standard curve can then be used to accurately determine the concentration of VEGF165 in any sample tested.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
VEGF 165A and VEGF 165b both isoforms of Vascular Endothelial Growth Factor (VEGF) play important roles in angiogenesis and vascular permeability. These proteins are part of the VEGF family and are derived from alternative splicing of the VEGFA gene. The mass of VEGF 165 isoforms is approximately 19 kDa. VEGF 165A serves as a potent pro-angiogenic factor while VEGF 165b acts as an anti-angiogenic counterpart. Expression of these proteins occurs mainly in endothelial cells but is also found in many tissues including lung heart and kidneys.
Biological function summary
VEGF 165 isoforms engage in modulating blood vessel formation and function. They achieve their roles by binding to and activating receptor tyrosine kinases namely VEGFR-1 and VEGFR-2 forming a ligand-receptor complex. This interaction sets off a cascade of downstream signaling events central to endothelial cell proliferation and migration. VEGF 165b despite binding the same receptors prevents these signals counteracting the function of VEGF 165A.
Pathways
VEGF 165 isoforms are significant parts of the VEGF signaling pathway and PI3K-Akt pathway. These pathways are central to cell survival growth and angiogenesis. In these pathways proteins such as PLCγ AKT and eNOS associate with VEGF 165 isoforms to mediate various physiological responses. This association highlights their roles in influencing cellular events related to vascular dynamics and homeostasis.
Abnormalities in VEGF 165 isoform expression relate to cancer and diabetic retinopathy. In many cancers overexpression of VEGF 165A facilitates tumor growth by promoting blood vessel formation. Conversely VEGF 165b is often diminished in such cases. In diabetic retinopathy imbalances between these isoforms exacerbate pathological angiogenesis leading to vision loss. Other proteins such as HIF-1α and PDGF-BB also connect with these isoforms in disease states impacting pathogenesis and offering potential therapeutic targets.