Description
Pervanadate is a protein tyrosine phosphatase inhibitor that is commonly used to increase tyrosine phosphorylation in cells. When cells are treated with pervanadate for 30 minutes they undergo significant tyrosine phosphorylation, as shown by western blotting using anti-Phosphotyrosine (PP2221).Fibroblast from rabbit spleen were harvested to generate a primary fibroblast cell line. Confluent cultures of rabbit fibroblasts were serum starved 2 hrs, then either left untreated or treated with pervanadate (1mM) for 30 minutes at 37 degree C. Cells were lysed in 1% SDS, 1.0mM sodium ortho-vanadate, 10mM Tris (pH7.4) buffer. Protein concentration was determined using the BCA method (Pierce) before diluting to final concentration and buffer