Quick Human VEGF ELISA Kit

Catalog Number:

S10E5

Background:

Vascular endothelial growth factor (VEGF) is one of the key players in angiogenesis, the process of new blood vessel formation. Blocking the interaction between VEGF and its receptors is one of many approaches to reduce tumor growth.

Applications:

This kit is developed to quickly titrate human VEGF sample in high throughput. It is for research use only.

Description:

These kits were validated using human VEGF protein (see figure). The detection linear range is 0.25 – 15.8 µg/ml; Sensitivity is 0.1 µg /ml.

Biosafety Level:

BSL-1

Organism:

Vascular endothelial growth factor (VEGF)

Assay Principles (Take neutralizing antibody as example):

This kit is developed for a sandwich ELISA format to specifically capture human VEGF protein in serum, plasma, tissue homogenates, cell culture supernatant and other biological fluids. The whole process should take about 2.5 hours. An antibody specific for VEGF has been pre-coated onto a detachable 96-well microtiter plate. The standards and test samples are added into the wells and the VEGF present in each sample is bound to the wells by the immobilized antibody. Following incubation, the wells are washed and then incubated with HRP labeled detector antibody, which binds the captured VEGF present in each well. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-colored product that changes to yellow after addition of stop solution. The density of yellow is proportional to the amount of VEGF captured in each well. The concentration of VEGF can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.

The kit consists of detachable 96-well plate pre-coated with VEGF-specific antibody, VEGF protein standard, HRP labeled detector antibody, TMB and sample dilution/wash/stop buffer.

Brief protocol (detailed protocol is provided with the product):

1. Add your samples containing human VEGF and provided purified VEGF protein standard into pre-coated plate (or strips based on sample size). Incubate at 37°C for 1 hour.
2. Wash wells. Add HRP labeled detector antibody to wells. Incubate at room temperature for 1 hour.
3. Wash wells. Add TMB and stop solution. Read absorbance (OD) at 450 nm. The OD Value reflects the amount of VEGF in the samples, which is then quantified against VEGF standard curve to calculate titer.

Storage and shipping condition:

Shipment with room temperature. The unopened kit is stable for 6 months from the date of manufacture when stored at 2°C to 8°C. Avoid light. The shelf life is 60 days from the date of opening.