Western blot analysis of Human VEGF was performed by loading 0. 5µg of VEGF transfected 293T cell culture supernatant and recombinant human VEGF onto a 4-12% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 3% BSA/TBST for at least 1 hour. The membrane was probed with a human VEGF polyclonal antibody (Product # P802) at a dilution of 1µg/ml overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20 and probed with a goat anti- rabbit-HRP secondary antibody (Product # 31460) at a dilution of 1:15,000 for at least one hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product #34075).
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Storage buffer||PBS with 4% BSA|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.125 - 0.5µg/ml|
|Western Blot (WB)||1 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
The VEGF-A monoclonal antibody clone # 16F1 (Product # M808) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody P802B (biotinylated conjugate of Product # P802).
Typical dilutions for sandwich ELISA include 1µg/ml for coating and 0.125-0.5µg/ml for detection.
The VEGF gene is a member of the PDGF/VEGF growth factor family and encodes a protein that is often found as a disulfide linked homodimer. This protein is a glycosylated mitogen that specifically acts on endothelial cells and has various effects, including mediating increased vascular permeability, inducing angiogenesis, vasculogenesis and endothelial cell growth, promoting cell migration, and inhibiting apoptosis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.