Pervanadate is a protein tyrosine phosphatase inhibitor that is commonly used to increase tyrosine phosphorylation in cells. Human umbilical vein endothelial cells treated with pervanadate for 30 minutes display a significant increase in tyrosine phosphorylation as shown by western blotting using anti-Phosphotyrosine.
Confluent cultures of human umbilical cord endothelial cells were serum starved 2 hrs at 37°C. Cells were then either left untreated (Cat.# EL7201) or treated with pervanadate (1 mM) for 30 minutes at 37°C (Cat # EL7301). Cells were lysed in 1% SDS, 1.0 mM sodium ortho-vanadate, 10 mM Tris (pH 7.4) buffer. Protein concentration was determined using the BCA method (Pierce) before diluting to final concentration and buffer.
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*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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