Bethyl Laboratories, Inc.

Bethyl Laboratories, Inc.

Antibodies to Kinases in the MAPK Pathway

April 7, 2010

Overview

The mitogen-activated protein kinase (MAPK) pathway can be activated by an assortment of extracellular stimuli such as growth factors, cytokines, and stress signals. Pathway activation leads to the coordination of a wide variety of cellular responses involving gene expression, proliferation, survival, apoptosis, metabolism, and differentiation. In mammals, there are five distinct groups of MAPKs: the extracellular signal-regulated kinases (ERKs), the c-Jun amino-terminal kinases (JNKs), the p38 isoforms, ERKs 3 and 4, and ERK5. In response to extracellular stimuli, ERKs, JNKs, and the p38 isoforms are activated by upstream MAPK kinases (MAPKKs) which are themselves activated by MAPKK kinases (MAPKKKs). For example ERK1 and ERK2 (ERK1/2) are activated by the upstream MAPKKs, MEK1 and MEK2 (MEK1/2). The MAPKKKs for MEK1/2 include A-Raf, B-Raf, Raf-1, and c-Mos which typically receive stimuli from growth factors via cell surface receptors. On the other hand, the p38 isoforms are activated by the MAPKKs, MEK3 and MEK6, which are mainly activated by environmental stress and inflammatory cytokine signals. The major substrates of MAPKs include MAPK-activated kinases (MKs) and transcription factors such as c-Jun, c-Myc, Max, CREB, p53, and ATF-2. The diverse biological functions influenced by MAPK signaling indicate that the MAPK network is complex. This complexity leaves volumes that require further study for a full understanding of the control of the MAPK modules as well as module cross-talk.

Selected Review
1. P. P. Roux and J. Blenis, "ERK and P38 MAPK-Activated Protein Kinases: a Family of Protein Kinases With Diverse Biological Functions," Microbiol.Mol Biol Rev. 68, no. 2 (2004): 320-344.

Bethyl Laboratories' Portfolio
AKT1 AKT2 ERK1 ERK2 HPK1 JIK/TAOK3 MAP4K4/HGK MEK1 MEK2 MEKK1 MEKK2 MST1,2/STK3,4 MST1/STK4 P38 MAPK PAK1 PAK2 PKC-alpha RAF1/c-RAF RSK1 RSK2	Detection of Human and Mouse p38 MAPK by Western Blot (h&m) and Immunoprecipitation (h). Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded), 293T (T; 50 mcg), and mouse NIH3T3 (M; 50 mcg) cells. Antibodies: Affinity purified rabbit anti-p38 MAPK antibody BL2225 (Cat. No. A300-707A) used for WB at 0.04 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 3 mcg/mg lysate (B). p38 MAPK was also immunoprecipitated using rabbit anti-p38 MAPK antibody BL2226 (Cat. No. A300-708A) at 3 mcg/mg lysate. For blotting immunoprecipitated p38 MAPK, ReliaBLOT^� Reagents and Procedures (Cat. No. WB120) were used. Detection: Chemiluminescence with exposure times of 30 seconds (A and B).	Detection of Human AKT2 by Western Blot and Immunoprecipitation. Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded) and 293T (T; 50 mcg) cells. Antibodies: Affinity purified rabbit anti-AKT2 antibody A302-209A used for WB at 0.04 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 10 mcg/mg lysate. AKT2 was also immunoprecipitated by rabbit anti-AKT2 antibody BL2163, which recognizes an upstream epitope. For blotting immunoprecipitated AKT2, ReliaBLOT^� Reagents and Procedures (Cat. No. WB120) were used. Detection: Chemiluminescence with exposure times of 30 seconds (A) and 3 seconds (B).

Search for Antibodies by Research Area
Bethyl Laboratories' portfolio of polyclonal antibodies commercially available includes over 5000 polyclonal antibodies to over 1500 proteins. We test our antibodies in a wide range of applications including ELISA, western blot, immunoprecipitation, immunohistochemistry, and immunocytochemistry. We have recently validated many of our antibodies for flow cytometry and ChIP. Our antibody range also includes secondary antibodies, loading controls and antibodies to epitope tags.

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