P38α sc 535

Manufactured by Santa Cruz Biotechnology

Sourced in United States

P38α (sc-535) is a primary antibody that targets the p38α mitogen-activated protein kinase (MAPK) protein. P38α is a key regulator of cellular responses to various environmental stresses and inflammatory cytokines. This antibody can be used for the detection of p38α in various applications, such as Western blotting, immunoprecipitation, and immunohistochemistry.

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Lab products found in correlation

Nfatc1 antibody, by BD (2 mentions) Blimp1 sc 47732, by BD (2 mentions) C fos sc 52, by Santa Cruz Biotechnology (2 mentions) Immobilon p membrane, by Merck Group (2 mentions) Western lightning plus ecl, by PerkinElmer (2 mentions) Sc409, by Merck Group (2 mentions) A20 tnfaip3, by Cell Signaling Technology (1 mentions) Irf8 sc 6058, by Santa Cruz Biotechnology (1 mentions) Actin, by MP Biomedicals (1 mentions) Goat anti mouse, by Jackson ImmunoResearch (1 mentions) Phospho p38 map kinase thr180 tyr182, by Cell Signaling Technology (1 mentions) Goat anti rabbit, by Jackson ImmunoResearch (1 mentions) Westernbright ecl, by Advansta (1 mentions) Cd16 cd32 2.4g2, by BD (1 mentions) Cd25 pc61, by Thermo Fisher Scientific (1 mentions) Hamster igg, by MP Biomedicals (1 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Cd3 145 2c11, by Thermo Fisher Scientific (1 mentions) Foxp3, by Thermo Fisher Scientific (1 mentions)

8 protocols using p38α sc 535

Western Blotting: A20, PLC γ2, p38α

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Western blotting was performed using standard methodology with the following antibodies: A20/TNFAIP3 (5630, Cell Signaling Technology) with 1:1000 dilution; PLC γ2 (sc-407, Santa Cruz Biotechnology) with 1:1000 dilution; and p38α (SC-535, Santa Cruz Biotechnology) with 1:1000 dilution.

Sokhi U.K., Liber M.P., Frye L., Park S., Kang K., Pannellini T., Zhao B., Norinsky R., Ivashkiv L.B, & Gong S. (2018). Dissection and function of autoimmunity-associated TNFAIP3 (A20) gene enhancers in humanized mouse models. Nature Communications, 9, 658.

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Western Blot Analysis of Transcription Factors

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Total cellular extracts were obtained using lysis buffer containing 150 mM Tris-HCl (pH 6.8), 6% SDS, 30% glycerol, and 0.03% Bromophenol Blue; 10% 2-ME was added immediately before harvesting cells. Cell lysates were fractionated on 7.5% SDS-PAGE, transferred to Immobilon-P membranes (Millipore), and incubated with specific antibodies. Western Lightning plus-ECL (PerkinElmer) was used for detection. NFATc1 antibody (556602, 1:1000) was from BD Biosciences; Blimp1 (sc-47732, 1:1000), c-Fos (sc-52, 1:1000), IRF8 (sc-6058, 1:1000), and p38α (sc-535, 1:3000), B-Myb (sc-390198, 1:1000) antibodies were from Santa Cruz Biotechnology; IRF1 (8478, 1:1000) antibody was obtained from Cell Signaling Technology.

Xia Y., Inoue K., Du Y., Baker S.J., Reddy E.P., Greenblatt M.B, & Zhao B. (2022). TGFβ reprograms TNF stimulation of macrophages towards a non-canonical pathway driving inflammatory osteoclastogenesis. Nature Communications, 13, 3920.

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Western Blot Antibody Detection Protocol

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Western blot analyses were performed with the following antibodies: anti-HA (hybridoma clone 12CA5, ATCC); p38α (sc-535) and p38β (sc-390984) (Santa Cruz Biotechnology, Santa Cruz, CA, USA); phospho-p38 MAP kinase (Thr180/Tyr182) (Cell Signaling, Danvers, MA, USA); actin (MP Biomedicals, Irvine, CA, USA). The immune detection was performed by peroxidase-conjugated secondary antibodies (goat anti-mouse; goat anti-rabbit; Jackson Immunoresearch, West Grove, PA, USA), and signals were developed using WesternBright ECL (Advansta, San Jose, CA, USA).

Melamed Kadosh D., Beenstock J., Engelberg D, & Admon A. (2023). Differential Modulation of the Phosphoproteome by the MAP Kinases Isoforms p38α and p38β. International Journal of Molecular Sciences, 24(15), 12442.

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Immune Cell Characterization Protocol

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DNFB, brefeldin A, and PF3644022 were from Sigma-Aldrich; SC409, PD98059, SP600125, and rapamycin from EMD Millipore; CFSE from Life Technologies; IFN-γ, IL-2, IL-4, IL-6, and TGF-β from Peprotech. Fluorescent-conjugated antibodies against the following markers were used in flow cytometry: CD3 (145–2C11), CD4 (GK1.5 and RM4–5), CD8 (53–6.7), CD25 (PC61.5), Foxp3 (anti-mouse/rat staining kit), IFN-γ (XMG1.2), IL-4 (BVD6–24G2), IL-17A (eBio17B7; all from eBioscience). Antibodies against the following proteins were used in cell stimulation, Fc receptor blocking, and cytokine neutralization: CD3 (145–2C11), CD28 (37.51), IFN-γ (XMG1.2), IL-12 (C17.8), IL-4 (11B11), CD16/CD32 (2.4G2; all from BD Pharmingen); and hamster IgG (MP Biomedicals). Antibodies against the following proteins were used in immunoblotting: ERK (9102), phosphorylated (p-) p38 (9211), p-JNK (9251), p-ERK (9101), p-S6K1 (9205), p-S6 (4858), p-MK2 (3007), p-TSC2 (3616; all from Cell Signaling Technology); p38α (sc-535; Santa Cruz Biotechnology); p38β (33–8700; Life Technologies); p38γ and p38δ (University of Dundee); JNK (554285, BD Pharmingen); Foxp3 (14–5773, eBioscience); and actin (A4700; Sigma-Aldrich).

Hayakawa M., Hayakawa H., Petrova T., Ritprajak P., Sutavani R.V., Jiménez-Andrade G.Y., Sano Y., Choo M.K., Seavitt J., Venigalla R.K., Otsu K., Georgopoulos K., Arthur J.S, & Park J.M. (2016). Loss of Functionally Redundant p38 Isoforms in T Cells Enhances Regulatory T Cell Induction. The Journal of Biological Chemistry, 292(5), 1762-1772.

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Protein Extraction and Western Blot Analysis

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Total cellular extracts were obtained using lysis buffer containing 150 mM Tris-HCl (pH 6.8), 6% SDS, 30% glycerol, and 0.03% Bromophenol Blue, with 10% 2-Mercaptoethanol added immediately before harvesting cells. Cell lysates were fractionated on 7.5% SDS-PAGE, transferred to Immobilon-P membranes (0.45 μm, Millipore), and incubated with specific antibodies. Western Lightning Plus-ECL (PerkinElmer) was used for detection. β-catenin antibody (9562, 1:1000) and Jag1 antibody (70109, 1:1000) were obtained from Cell Signaling Technology. Nfatc1 antibody (556602, 1:1000) was obtained from BD Biosciences; Blimp1 (sc-47732, 1:1000), c-Fos (sc-52, 1:1000), OPG/Osteoprotegerin (sc-390518, 1:1000) and p38α (sc-535, 1:3000) antibodies were purchased from Santa Cruz Biotechnology.

Qin Y., Shirakawa J., Xu C., Chen R., Ng C., Nakano S., Elguindy M., Deng Z., Prasanth K.V., Eissmann M.F., Nakagawa S., Ricci W.M, & Zhao B. (2024). Malat1 fine-tunes bone homeostasis by orchestrating cellular crosstalk and the β-catenin-OPG/Jagged1 pathway. Research Square.

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Inflammatory Signaling Pathway Protocol

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LPS from Escherichia coli 0111:B4 (L2630), LPS from Salmonella enterica serotype minnesota Re 595 (L9764), peptidoglycan (79682), and actin antibody (A2228) were purchased from Sigma (St. Louis, MO, USA). Antibodies for phospho-PKC-α (sc-12356), phospho-PKC-δ (sc-11770), PKC (sc-80), phospho-IκB-α (sc-8404), IκB-α (sc-371), iNOS (sc-651), COX-2 (sc-1745), ERK1/2 (sc-135990), JNK1 (sc-1648) and p38α (sc-535) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibody for NLRP3 (AG-20B-0006-C100) was purchased from Adipogen (San Diego, CA, USA). Mouse IL-1β (88-7013-88), mouse IL-6 (88-7064-88), mouse TNF-α (88-7324-88), human IL-6 (88-7066-88) and human TNF-α (88-7346-88) ELISA kits were purchased from eBioscience (San Diego, CA, USA). MAPK family antibody sampler kit (9910T) was purchased from Cell Signaling Technology, Inc (Danvers, MA, USA). Pam3CSK4 (tlrl-pms), ATP (tlrl-atp), nigericin (tlrl-nig) and monosodium urate crystal (MSU) (tlrl-msu) were purchased from InvivoGen (San Diego, CA, USA).

Li L.H., Ju T.C., Hsieh C.Y., Dong W.C., Chen W.T., Hua K.F, & Chen W.J. (2017). A synthetic cationic antimicrobial peptide inhibits inflammatory response and the NLRP3 inflammasome by neutralizing LPS and ATP. PLoS ONE, 12(7), e0182057.

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Comprehensive Immunoblotting and Immunostaining Protocols

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Unless otherwise specified all chemicals were from Sigma. Antibodies were from Biolegend: HA (MMS-101P), K5 (PRB-160P), K6 (PRB-169P), Loricrin (PRB-145P); Santa Cruz Biotechnology: p38α (sc-535), p63 (sc-8431), p65-NF-κB (sc-372), ERK (sc-154), GR (sc-1004), HSD11B2 (sc-20176), IκB(sc-371), Mkp1/2 (sc-1102), MR (sc-11412, ChIP), LaminA (sc-6214); Abcam: MR (ab64457); Cell Signalling Technology: p-ERK (Thr202/Tyr204; #4376), p-p38 (#9211S and #4631 for immunohistochemistry), p-JNK (#9251), JNK (#9252); ThermoFisher: GFP (A6455); Sigma: Tubulin (T6199), Actin (A2066); Roche: BrdU (11170376001) and BD Biosciences: E-cadherin (610181), Ly6G (551459). Secondary biotin-conjugated anti-rabbit, anti-mouse and anti-rat antibodies (Jackson ImmunoResearch) and secondary Alexa Fluor® anti-rabbit (555, A-31572) antibody (ThermoFisher) were used for immunostaining. Secondary peroxidase-conjugated anti-rabbit (NA934), anti-mouse (NA931) antibodies (GE Healthcare), and anti-goat (Jackson Immunoresearch) were used for immunoblotting.

Bigas J., Sevilla L.M., Carceller E., Boix J, & Pérez P. (2018). Epidermal glucocorticoid and mineralocorticoid receptors act cooperatively to regulate epidermal development and counteract skin inflammation. Cell Death & Disease, 9(6), 588.

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Analyzing p38 and p63 Signaling

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DMBA, TPA, 5-bromo-2’-deoxyuridine (BrdU), cycloheximide, 5Z-7-Oxozeaenol, and verapamil were from Sigma-Aldrich; MG132, anisomycin, SB202190, and SC409 from EMD Millipore; D-JNKi from BML; and murine IL-1α from R&D systems. Antibodies against the following proteins were used in immunoblotting after 1:1000 dilution: p38α (sc-535) and p63 4A4 (sc-8431; both from Santa Cruz Biotechnology); p-p38 (9211), p-MK2 (3007), and p-Ser66/68-p63 (4981; all from Cell Signaling Technology); p-Ser301-p63 (PA5–39827) and p-Ser361-p63 (PA5–38380; both from Thermo Fisher Scientific); and actin (A4700; Sigma-Aldrich). Antibodies specific to the following proteins were used in immunofluorescence staining after 1:50 to 1:1000 dilution: p38α (sc-535), p63 H-137 (sc-8343), and p63 4A4 (sc-8431; all from Santa Cruz Biotechnology); p-p38 (9211) and p-Ser66/68-p63 (4981; both from Cell Signaling Technology); Ki67 (ab16667) and MMP13 (ab39012; both from Abcam); KRT14 (PRB-155P) and KRT15 (PCK-153P; both from Covance); CD3e (SP7) and KRT1 (ab9286; both from Abcam); and Ly6G/Gr-1 (553125; BD Biosciences). Hoechst 33324 and antibodies conjugated with Alexa Fluor 488 or Alexa Fluor 595 were from Molecular Probes.

Choo M.K., Kraft S., Missero C, & Park J.M. (2018). The protein kinase p38α destabilizes p63 to limit epidermal stem cell frequency and tumorigenic potential. Science signaling, 11(551), eaau0727.

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