MAP Kinase p44/42 (Thr202/Tyr204) BioAssay(TM) Phosphorylation Detection Kit

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.\n\nApplications: \nSuitable for use in Western Blotting, Immunoprecipitation, Immunohistochemistry, Immunocytochemistry. Other applications not tested.\n\nRecommended Dilution:\nWestern blotting: 1:1000 1:1000\nImmunoprecipitation: 1:100 1:50\nImmunocytochemistry: 1:250\n\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay 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.

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SPECIFICATIONS

Catalog Number

M2352-32

Size

1Kit

Applications

ICC, IHC, IP, WB

Purity

Purified by protein A and peptide affinity chromatography.

References

1. Marshall, C.J. (1995) Cell 80, 179

Additional Info

Phospho-p44/42 MAP Kinase (Thr202/Tyr204) Antibody detects phosphorylated threonine 202 and tyrosine 204 of p44 and p42 MAP Kinases (Erk1 and Erk2) but does not appreciably crossreact with the corresponding phosphorylated threonine and tyrosine of either JNK/SAPK or p38 MAP Kinase homologues.