Western blot analysis of extracts from PAE cells transfected with either a mock expression vector or human VEGFR-1, using VEGF Receptor 1 Antibody.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
VEGF Receptor 1 Antibody detects endogenous levels of VEGFR-1. This antibody does not cross-react with VEGFR-2 and -3 nor does it detect the sVEGFR-1 variant.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr1307 of human VEGFR-1. Antibodies are purified by protein A and peptide affinity chromatography.
The vascular endothelial growth factor (VEGF) receptor (VEGFR-1, Flt-1) is a 180 kDa receptor tyrosine kinase belonging to the VEGFR (Flt) family (1-3). The receptor is comprised of seven extracellular Ig-like domains, a single transmembrane region and cytoplasmic tail containing the active kinase domain (1,2). VEGFR-1 plays an important role in endothelial cell function and normal vascular development, as well as in hematopoietic function (2,3). VEGF-A is a high affinity ligand of VEGFR-1. VEGFR-1 also binds VEGF-B and PLGF (2). Ligand binding results in very little VEGFR-1 kinase activation, and VEGFR-1/VEGF-A binding negatively regulates VEGF function by diverting the growth factor from other functional VEGF receptors (3).
Two forms of the VEGF receptor 1 are found in cells. Both the membrane-bound form described above and a soluble isoform of VEGFR-1 (sVEGFR-1 or sFlt-1) bind the VEGF ligand with high affinity (4). Full-length VEGFR-1 and the truncated, soluble protein are encoded by the same gene and are generated through differential splicing. Both proteins are associated with an array of human disorders and are potential candidates for therapeutic study (5).
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