The progression of cells through the cell cycle is regulated by a family of proteins designated cyclin-dependent kinases (Cdks). Sequential activation of individual members of this family and their consequent phosphorylation of critical substrates promotes orderly progression through the cell cycle. Multiple proteins are encoded by the tumor suppressor gene CDKN2A (MTS1/ p16INK4a) via translation of alternate reading frames, resulting in the pro- duction of the p19 ARF protein in mice and the p14 ARF protein in humans. p14 ARF induces an increase in MDM2 and p21 levels and leads to cell cycle arrest in both G1 and G2/M. p14 ARF is negatively regulated by p53 and is known to bind directly to MDM2. CDKN2A also encodes the mitotic protein p16, which binds to and inhibits the Cdk4/cyclin D complex.
Flow Cytometry (1-2ug/million cells), Immunofluorescence (1-2ug/ml), Western Blot (1-2ug/ml), Immunohistochemistry (Formalin-fixed) (1-2ug/ml for 30 min at RT)(Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA,
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