MAP Kinase Signaling
p38α MAP Kinase #7474
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Source / Purification
The GST-Kinase fusion protein was produced using a baculovirus expression system with a construct expressing full length human p38α MAP Kinase (Met1-Ser360) (GenBank Accession No. NM_139012) with an amino-terminal GST tag. The protein was activated by co-expressing with active MKK3 and purified by one-step affinity chromatography using glutathione-agarose.
Description
Purified recombinant full length human p38α MAP Kinase (Met1-Ser360) kinase, supplied as a GST fusion protein.
Quality Control
The theoretical molecular weight of the GST-p38α MAP Kinase fusion protein is 68 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. p38alpha MAP kinase activity was determined using a radiometric assay [Fig.2].
Gel Staining
Figure 1. The purity of the GST-p38α MAP Kinase fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. p38α MAP Kinase activity was measured in a radiometric assay using the following reaction conditions: 4 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 0.4 mM EDTA, 4 mM MgCl2, 0.05 mM DTT, 40 ng/μl BSA, 50 μM ATP, Substrate: ATF2 250 ng/μL and recombinant p38α MAP Kinase: variable.
Background
p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAP kinase, p38α, β, γ (also known as ERK6 or SAPK3) and δ (also known as SAPK4) have been identified. Similar to the SAPK/JNK pathway, p38 MAP kinase is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharides (LPS), UV light and growth factors (1-5). MKK3, MKK6 and SEK activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182. Activated p38 MAP kinase has been shown to phosphorylate and activate MAPKAP kinase 2 (3) and to phosphorylate the transcription factors ATF-2 (5), Max (6) and MEF2 (5-8).
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- Yang, S.H. et al. (1999) Mol. Cell. Biol.19, 4028 - 4038.
Application References
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Companion Products
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
