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.
Confluent cultures of A431 cells were serum starved overnight. Cells were then either left untreated (Cat.# AL9401) or treated with pervanadate (1 mM) for 30 minutes at 37°C (cat.# AL9501). 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.
Western blot analysis of A431, HeLa, and Jurkat cells (20 µg/lane) untreated or treated with pervanadate (1 mM) for 30 min. Blots were probed with anti-Phosphotyrosine rabbit polyclonal.
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*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blotmobilities of known proteins with similar MW.
This kit contains:
