Both p44 and p42 MAP kinases (Erk1 and Erk2) function in a protein kinase cascade that plays a critical role in the regulation of cell growth and differentiation (1-4). MAP kinases are activated by a wide variety of extracellular signals including growth and neurotrophic factors, cytokines, hormones, and neurotransmitters. Activation of MAP kinases occurs through phosphorylation of threonine and tyrosine (202 and 204 of human MAP kinase [Erk1]) at the sequence T*EY* by a single upstream MAP kinase kinase (MEK) (figure 1) (5,6). Both kinases are known to weakly autophosphorylate on tyrosine (5). The p44/42 MAP Kinase Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of p44 and p42 MAP Kinases (Erk1 and Erk2). The kit includes a phospho-MAP kinase antibody that recognizes threonine 202 and tyrosine 204 phosphorylated MAP kinase, a control MAP kinase antibody (phosphorylation-state independent), protein controls for western blots and the HRP Detection System. Figure 1: Growth factor and cytokine activated MAP kinase signaling pathways. The activation sites of the three different MAP kinase families, ERK, SAPK and p38/RK are shown as are the activated transcription factors.
Suitable for use in Western Blotting, Immunoprecipitation, Immunohistochemistry, Immunocytochemistry. Other applications not tested.
Western blotting: 1:1000 1:1000
Immunoprecipitation: 1:100 1:50
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20 degrees C or colder. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.