P38mapk d13e1

Manufactured by Cell Signaling Technology

Sourced in United States

P38MAPK (D13E1) is a rabbit monoclonal antibody that recognizes p38 mitogen-activated protein kinase (MAPK). p38 MAPK is a serine/threonine protein kinase that is involved in the cellular response to various stimuli, such as stress, cytokines, and growth factors.

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Lab products found in correlation

Sapk jnk, by Cell Signaling Technology (1 mentions) Phospho sapk jnk, by Cell Signaling Technology (1 mentions) Hrp conjugated anti rabbit antibody, by Bio-Rad (1 mentions) Protein assay dye, by Bio-Rad (1 mentions) Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Phosphatase inhibitor cocktail, by Cell Signaling Technology (1 mentions) Leupeptin, by Merck Group (1 mentions) Aprotinin, by Merck Group (1 mentions) Phospho p38 map kinase, by Cell Signaling Technology (1 mentions) Ab109235, by Abcam (1 mentions) Jak2 d2e12, by Cell Signaling Technology (1 mentions) Pi3 kinase p85 19h8, by Cell Signaling Technology (1 mentions) Phosstop phosphatase inhibitor, by Roche (1 mentions) Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions) Tmt kit, by Thermo Fisher Scientific (1 mentions) Acetonitrile, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Lonsera (1 mentions) Trypsin edta, by Beyotime (1 mentions) Bca protein assay kit, by Beyotime (1 mentions)

5 protocols using p38mapk d13e1

Western Blot Analysis of ABC Transporters

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Cells were washed twice with ice-cold PBS, harvested by scraping and whole cell lysates were prepared in RIPA buffer containing phosphatase and protease inhibitors (#SC-24949; Santa Cruz Biotechnologies). Cell lysates were incubated on ice for 30 min and clarified by centrifugation at 20,000 g for 30 min. Protein content was estimated using Bio-Rad protein Assay Dye (Catalog#500–0006). Equal amounts of protein (40 μg) were resolved in 12% SDS-PAGE gels, blotted onto PVDF membranes (#IPVH00010; EMD Millipore) and probed with antibodies for MDR1(D3H1Q; 1:1000; Cell Signaling), MRP1(D708N; 1:1000; Cell Signaling), ABCG2 (D5V2K; 1:1000; Cell Signaling), phospho-p38MAPK (D3F9; 1:1000; Cell Signaling), p38MAPK (D13E1; 1:1000; Cell Signaling), phospho-p44/42 MAPK (Erk1/2) (D13.14.4E; 1:1000; Cell Signaling), p44/42 MAPK (Erk1/2) (137F5; 1:1000; Cell Signaling) phospho-SAPK-JNK (81E11;1:1000; Cell Signaling), SAPK-JNK (#9252; 1:1000; Cell Signaling) and GAPDH (14C10; 1:1000; Cell Signaling). Secondary HRP-conjugated anti-rabbit antibody (#170–6515; 1:1000; Bio-Rad) was used. The antigen-antibody complexes were detected by chemiluminescence using Super Signal West Pico Chemiluminescent Substrate (#34080; Thermo Scientific, USA). Densitometry was performed using Image J software (NIH) and histograms showing relative expression of MDR1, MRP1 and ABCG2 normalized to GAPDH were generated.

Sonowal H., Pal P.B., Wen J.J., Awasthi S., Ramana K.V, & Srivastava S.K. (2017). Aldose reductase inhibitor increases doxorubicin-sensitivity of colon cancer cells and decreases cardiotoxicity. Scientific Reports, 7, 3182.

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Analyzing Protein Signaling Cascades

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The CMECs were homogenized in RIPA lysis buffer (#P0013C; Beyotime) containing 1X Phosphatase Inhibitor Cocktail (#5870S; Cell Signaling Technology, Beverly, MA, USA) and 1 µg/ml each of aprotinin (A1153; Sigma-Aldrich) and leupeptin (#L2884; Sigma-Aldrich). Forty micrograms of protein was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to polyvinylidene difluoride (PVDF) membranes, and then probed with antibodies for XO (#ab109235; Abcam), phospho-PI3 kinase p85 (Tyr458)/p55 (Tyr199) (#4228), PI3 kinase p85 (19H8) (#4257), phospho-p38 MAP kinase (Thr180/Thr182) (#4631), p38 MAPK (D13E1) (#8690), JAK2 (D2E12) (#3230) and phospho-JAK2 (Tyr1007/1008) (C80C3) (#3776) (all from Cell Signaling Technology, Beverly, MA, USA). The same membranes were reprobed with an antibody for tubulin (#AT819; Beyotime). The blotting film was quantified using a scanner and a densitometry program (ImageJ; https://imagej.nih.gov/ij/index.html) (24 ). To quantify the phosphor-specific signal in the activated samples, the background was subtracted and the band was normalized to the amount of tubulin or total target protein in the lysate.

Zhang Y.Q., Hu S.Y., Chen Y.D., Guo M.Z, & Wang S. (2016). Hepatocyte growth factor inhibits hypoxia/reoxygenation-induced activation of xanthine oxidase in endothelial cells through the JAK2 signaling pathway. International Journal of Molecular Medicine, 38(4), 1055-1062.

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Activation of p38 MAPK Pathway

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Reagents: Dulbecco's modified eagle's medium (DMEM) (Sigma-Aldrich, MO, USA); fetal bovine serum (FBS) (Lonsera, UY, South America); p38 mitogen activated protein kinase (p38 MAPK) (D13E1) (8690, Cell Signaling Technology, MA, USA); phospho-p38 MAPK (P-p38 MAPK) (Thr180/Tyr182) (4631, Cell Signaling Technology, MA, USA); GAPDH (AF7021, Affinity Biosciences, Beijing, China); horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (S0001, Affinity Biosciences, Beijing, China); phosSTOP phosphatase inhibitor (Roche, Basel, Switzerland); GC-rich PCR master mix (Sangon Biotech, Shanghai, China); acetonitrile (Fisher Chemical, Fairlawn, USA); triethylammonium bicarbonate (TEAB) (Sigma-Aldrich, MO, USA); Escherichia coli BL21(DE3) competent cells (Solarbio, Beijing, China); Ni NTA Beads 6FF (Smart-Lifesciences, Jiangsu, China); BCA protein assay kit (Beyotime, Shanghai, China); endofree plasmid midi kit (Cwbiotech, Beijing, China); fastpure gel DNA extraction mini kit (Vazyme Biotech, Jiangsu, China); Ni-NTA sensors (ForteBio, CA, USA); tandem mass tagging (TMT) Kit (Thermo, Waltham, USA). Ethylene diamine tetraacetic acid (EDTA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and trypsin-EDTA (TE) were obtained from Beyotime Technology (Shanghai, China).

Lu C., Fan L., Zhang P.F., Tao W.W., Yang C.B., Shang E.X., Chen F.Y., Che C.T., Cheng H.B., Duan J.A, & Zhao M. (2021). A novel P38α MAPK activator Bruceine A exhibits potent anti-pancreatic cancer activity. Computational and Structural Biotechnology Journal, 19, 3437-3450.

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PAHs and Cell Signaling Antibodies

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PAHs including phenanthrene (cat#73338), fluoranthene (cat#11474), benzo(a)pyrene (cat#51968), and benzo(b)fluoranthene (cat#30958) were purchased from Sigma-Aldrich (Darmstadt, Germany). The monoclonal rabbit cyclin D1 (92G2, cat#2978), phospho-p38 MAPK (Thr180/Tyr182) (D3F9, cat#4511), p38 MAPK (D13E1, cat#8690), p53 (D2H9O, cat#32532), and GAPDH (14C10, cat#2118) were obtained from Cell Signaling Technology (Beverly, MA, USA).

Pattarachotanant N., Prasansuklab A, & Tencomnao T. (2021). Momordica charantia L. Extract Protects Hippocampal Neuronal Cells against PAHs-Induced Neurotoxicity: Possible Active Constituents Include Stigmasterol and Vitamin E. Nutrients, 13(7), 2368.

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Immunoblot analysis of signaling proteins

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Equal amounts of whole cell lysates were separated on a 10–12% SDS-polyacrylamide gel. The blotting membranes were probed using the following antibodies: NDRG1 (42–6200; Invitrogen), SAPK/JNK (56G8, #9258, Cell Signaling, Danvers, MS, USA), phospho-SAPK/JNK (Thr183/Tyr185, 81E11, #8690, Cell Signaling), p44/42 MAPK (ERK 1/2; 137F5, #4695, Cell Signaling), phospho-p44/42 MAPK (ERK 1/2, Thr202/Tyr204, #9101, Cell Signaling), p38 MAPK (D13E1, #8690, Cell Signaling), phospho-p38 MAPK (Thr180/Tyr182, #9211, Cell Signaling), AMPKα1/2 (#5831, Cell signaling), phospho-AMPKα1/2 (Thr 172; #2535, Cell Signaling), GDF15 (Ab206414, Abcam, Cambridge, MA, USA), alpha-smooth muscle actin (α-SMA) (Ab5694, Abcam), uroplakin II (UPK2) (LS-C41569; LifeSpan BioSciences, Seattle, WA, USA), and β-actin (MAB1501, Merck Millipore, Burlington, MA, USA). Band intensities were detected by the Western lightning plus-ECL detection system (PerkinElmer Inc, Waltham, MA, USA), recorded using the LuminoGraph II (Atto Corporation, Tokyo, Japan), and analyzed using the GeneTool Program of the ChemiGenius (Syngene Cambridge, UK).

Hou C.P., Tsui K.H., Chang K.S., Sung H.C., Hsu S.Y., Lin Y.H., Yang P.S., Chen C.L., Feng T.H, & Juang H.H. (2021). Caffeic acid phenethyl ester inhibits the growth of bladder carcinoma cells by upregulating growth differentiation factor 15. Biomedical Journal, 45(5), 763-775.

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