Mitogen-Activated Protein Kinase 14 (MAPK14) (pThr180,pTyr182) antibody
Alternatives Western Blotting (WB), BioImaging (BI), Intracellular Flow Cytometry (ICFC)
|3 references available|
|Quantity||50 µg (250 µg/ml) (Variants)|
|Price||Product not available in this region.|
|Alternative name||p38 MAPK|
|Immunogen||Phosphorylated Human p38 MAPK (pT180/pY182) Peptide|
|Cross-Reactivity||Mouse (Murine), Rat (Rattus)|
Activation of the immune and inflammatory responses often involves the recognition of bacterial endotoxin (lipopolysaccharide or LPS). Binding of LPS by monocytes results in the production and release of proinflammatory cytokines, such as IL-1 and TNF. LPS-induced signaling cascades involve members of the Ser/Thr protein kinase family known as the Mitogen Activated Protein Kinases (MAPKs). MAPK signal transduction pathways mediate the effects of various extracellular stimuli on biological processes such as proliferation, differentiation, and death. The p38 MAPKs include p38alpha (MAPK14), beta (MAPK11), gamma (MAPK12), and delta (MAPK13). These Ser/Thr kinases are activated by dual phosphorylation on threonine (T) and tyrosine (Y) within the motif Thr-Gly-Tyr located in kinase subdomain VIII. Activation of p38 MAPK is mediated specifically by the MAP Kinase Kinases, MKK3, MKK4, and MKK6. This leads to the activation of multiple transcription factors (NF-kappaB, ATF-2, Elk-1, and CHOP) that induce expression of many different genes, including proinflammatory cytokine genes. Thus, p38 MAPKs are central kinases in multiple signal transduction pathways.
The 36/p38 (pT180/pY182) monoclonal antibody recognizes the conserved dual phosphorylated site pT180/pY182 of p38alpha, beta, gamma, and delta.
Synonyms: MK14, 11, 12, 13, CSBP1, SAPK2, 2A, 3, 4, MX12, ERK-6, ERK5
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.
|Molecular Weight||38-42 kDa|
Related Products: ABIN968535, ABIN968632, ABIN968769
|Synonyms||RK, p38, EXIP, Mxi2, CSBP1, CSBP2, CSPB1, PRKM14, PRKM15, SAPK2A, p38ALPHA, Hog, CRK1, CSBP, Exip, Csbp1, Csbp2, Prkm14, Prkm15, Sapk2A, p38Hog, p38alpha, MGC105413, p38b, zp38b, MAPK14, MGC142910, 186F5S, BG:DS00797.3, D-p38, D-p38b, Dm p38b, Dp38, ESTS:186F5S, Mpk34C, anon-sts23, p38 MAPK, p38 beta, p38B, p38beta, DmelCG7393, CG7393|
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 min. at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate 1 hr.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hr.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.
|Purification||Purified from tissue culture supernatant or ascites by affinity chromatography.|
|Buffer||Aqueous buffered solution containing BSA, glycerol.|
|Preservative||0.09% Sodium azide.|
|Storage||Store undiluted at -20° C.|
|Restrictions||For Research Use only|
Han, Lee, Bibbs et al.: "A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells." in: Science (New York, N.Y.), Vol. 265, Issue 5173, pp. 808-11, 1994 (PubMed).
Brunet, Pouysségur: "Identification of MAP kinase domains by redirecting stress signals into growth factor responses." in: Science (New York, N.Y.), Vol. 272, Issue 5268, pp. 1652-5, 1996 (PubMed).
Winston, Chan, Johnson et al.: "Activation of p38mapk, MKK3, and MKK4 by TNF-alpha in mouse bone marrow-derived macrophages." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 159, Issue 9, pp. 4491-7, 1997 (PubMed).