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Technical Data Sheet

PP3411

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p38α MAP Kinase (Tyr-323), phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms, Ck
38 kDa

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A) Western blot image of GST-recombinant p38 (K53M) mutant kinase untreated (lanes 1 & 3) or treated with Fyn kinase (lanes 2 & 4). B) Western blot analysis of p38 phosphorylation in mouse macrophages stimulated with 1 mM pervanadate for 30 min. (lanes 1 & 3) then the blot was treated with alkaline phosphatase (lanes 2 & 4). Both blots were probed with anti-p38α (a.a. 319-328) (lanes 1 & 2) or anti-p38α (Tyr-323) (lanes 3 & 4).

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Immunocytochemical labeling of p38 MAPK in pervanadate-treated mouse C2C12. The cells were labeled with mouse monoclonal p38α MAPK and rabbit polyclonal p38 MAPK (Tyr-323) antibodies, then the antibodies were detected using appropriate secondary antibodies conjugated to Cy3.

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Application
Dilution


ELISA
1:2000


ICC
1:300


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
Product References
Alam, M.S. et al. (2015) Nat Medicine 21:1337. WB, IF: human T-cells, CD3 activated
Lanna, A. et al. (2014) Nat Immunol. 15(10):965. WB: human T-cells, CD3 activated
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Background
p38 MAP kinase (MAPK), also called RK, CSBP, and SAPK2a, is the mammalian orthologue of the yeast HOG kinase. This family of kinases participates in signaling cascades that control cellular responses to cytokines and stress. Four isoforms of p38 MAPK (α,β,γ,δ) have been identified. Similar to the SAPK/JNK pathway, p38 MAPK is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharides, UV light, and growth factors. MKK3 and SEK activate p38 MAPK by dual phosphorylation at Thr-180/Tyr-182. Activated p38 MAPK has been shown to phosphorylate and activate MAPKAP kinase 2 and to phosphorylate the transcription factors ATF-2, Max and MEF2. T cells possess an alternative pathway for p38 activation where stimulation of the antigen receptor (TCR) induces phosphorylation of p38 on Tyr-323. This site is required for TCR-mediated phosphorylation of Thr-180 and catalytic activity. Thus, Tyr-323 may also have important roles in regulating p38 MAP kinase pathways.


Background References
Han, J. et al. (1994) Science 265:808.
Lee, J. C. et al. (1994) Nature 372:739.
Salvador, J.M. et al. (2005) Nat Immunol. 6(4):390.
Jirmanova, L. et al. (2009) Blood 113(10):2229.
Immunogen
Phospho-p38α MAP Kinase (Tyr-323) synthetic peptide (coupled to KLH) corresponding to amino acid residues surrounding tyrosine 323 in mouse p38α. This peptide sequence is highly conserved in human and rat p38α, and has high homology to the conserved site in p38β.
Buffer and Storage
Rabbit polyclonal, affinity-purified antibody is supplied in 100μl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
This antibody was cross-adsorbed to a phospho-tyrosine peptide before affinity purification using phospho-p38α (Tyr-323) peptide (without carrier). The antibody detects a 38 kDa* band corresponding to p38α on SDS-PAGE immunoblots of human Jurkat, K562, and mouse macrophage (J774A.1) cells treated with pervanadate. In addition, the antibody detects recombinant p38 MAP Kinase when treated with Fyn kinase (The in vitro kinase model was kindly provided by Dr. Paul Mittelstadt from the Laboratory of Immune Cell Biology at the NCI.)

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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