Western Blotting Experiment. Extracts prepared from CKR/PAE cells, which overexpress a chimeric protein containing the M-CSF receptor extracellular domain, the VEGFR2 transmembrane domain, and the VEGFR2 cytoplamic domain (lane 1; 1 µg total protein), or extracts prepared from HUVEC cells (lane 2; 20 µg total protein), were resolved by SDS-PAGE on an 8% polyacrylamide gel and transferred to PVDF. Membranes were blocked for one hour at room temperature in a 5% BSA-TBST buffer and incubated with the VEGFR2 pan antibody for two hours at room temperature in a 3% BSA-TBST buffer. After washing, membranes were incubated with goat F(ab and quote;)2 anti-rabbit IgG HRP conjugate (Cat. no. ALI4404) and bands were detected using the Pierce SuperSignal™ method. The figure presented at left demonstrates that this antibody reacts with the chimeric VEGFR2 expressed by CKR/PAE cells (mature form Mr=180 kDa, lane 1), as well as with the wild type VEGFR2 expressed by HUVEC cells (mature form Mr= 230 kDa, lane 2). A nonspecific band of varying intensity is observed at 100 kDa in all cell lines tested.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a synthetic peptide (KLH-coupled) corresponding to residues near the carboxy-terminal sequence of human VEGF Receptor 2.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* 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.
Flk-1/KDR vascular endothelial growth factor receptor 2, a cell membrane receptor kinase, is a high affinity receptor for vascular endothelial growth factor and is putatively involved in the growth of endothelial cells and angiogenesis. It contains seven immunoglobulin like sequences in the extracellular domain and exhibits sequence similarity to Flt-1 and Flt-4.
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