Rapamycin, sodium dichromate (Na2Cr2O7·H2O) [Cr(VI)], 2,3-diaminonaphthalene (DAN), mercury(II) chloride (HgCl2), sodium hydroxide (NaOH), NO inhibitor aminoguanidine (AG), p38 inhibitor SB203580, NO donor diethylamine NONOate sodium salt, Bafilomycin A1, Chloroquine diphosphate salt, and S-nitrosocysteine were obtained from Sigma–Aldrich (St. Louis, MO). The fluorogenic caspase-9 substrate, LEHD–amino-4-methylcoumarin (AMC), was from Alexis Biochemicals (San Diego, CA). Diaminofluorescein (DAF)-diacetate (DA) was purchased from Molecular Probes (Eugene, OR). Antibodies for Rabbit IgG, Bcl-2, p62, Beclin-1, Atg5, Atg12, P-p38/p38, β-actin, and peroxidase-labeled secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA). Lipofectamine 2000 was purchased from Life Technologies (Carlsbad, CA). The Bcl-2 inhibitor ABT-737 and a Bcl-2 antibody for immunoprecipitation were obtained from Santa Cruz Biotechnologies (Dallas, TX). pAb anti-LC3 antibody (HRP) was from Novus Biologicals (Littleton, CO).
P38 inhibitor sb203580
Manufactured by Merck Group
Sourced in United States, Germany
The P38 inhibitor SB203580 is a small-molecule compound that selectively inhibits the p38 mitogen-activated protein kinase (MAPK) signaling pathway. It is a commonly used research tool for studying the role of the p38 MAPK pathway in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Jnk inhibitor sp600125, by Merck Group (7 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Anti chchd4, by Proteintech (3 mentions)
Anti ha tag clone 3f10, by Roche (3 mentions)
Anti β actin and anti alpha tubulin, by Merck Group (3 mentions)
Restriction endonuclease, by New England Biolabs (3 mentions)
Anti tim23, by BD (3 mentions)
Pi3k inhibitor ly294002, by Merck Group (3 mentions)
Pd98059, by Merck Group (3 mentions)
P jnk, by Cell Signaling Technology (3 mentions)
Rapamycin, by Merck Group (2 mentions)
β actin, by Santa Cruz Biotechnology (2 mentions)
Cytochrome c, by Santa Cruz Biotechnology (2 mentions)
Bay 11 7092, by Merck Group (2 mentions)
Anti pgc 1α, by Santa Cruz Biotechnology (2 mentions)
Pka inhibitor h89, by Merck Group (2 mentions)
Anti hsp60, by Enzo Life Sciences (2 mentions)
Pkc inhibitor go6983, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Chloroquine diphosphate salt, by Merck Group (1 mentions)
20 protocols using p38 inhibitor sb203580
1
Mechanistic Insights into Chromium-Induced Autophagy
2
Investigating Anticancer Mechanisms using Cell Culture
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All culture materials were purchased from Gibco (Grand Island, NY, USA). 1-[4-(Furo[2,3-b]quinolin-4-ylamino)phenyl]ethanone (CIL-102), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), ROS scavenger (N-acetyl cysteine [NAC]), 2,7-dichlorodihydrofluorescein diacetate (H2DCFDA), dihydroethidium (DHE), ERK inhibitor (PD98059), c-Jun N-terminal kinase (JNK1/2) inhibitor (SP600125), p38 inhibitor (SB203580), and mTOR inhibitor (rapamycin) were purchased from Sigma (St. Louis, MO, USA). Mouse monoclonal antibodies against cyclin A, cyclin D1, cyclin E, cyclin B1, acetylation of H3 (Ac-Histone H3) at Lys 9 and Lys 14, cytochrome c, caspase-3, -8, -9 and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit polyclonal antibodies against cdk2, ERK1/2Thr202Tyr204, p38 Thr180Tyr182, and JNK1/2 Thr183Tyr185 mouse monoclonal cdc2, t-BID, p300, CBP, p21, GADD45, and Fas-L antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). SDS, NP-40, sodium deoxycholate, protease inhibitor cocktails were purchased from Sigma (St Louis, MO, USA).
3
B Cell Chemotaxis Assay in Transwells
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Mouse spleen cells were treated with ammonium chloride potassium buffer to lyse red blood cells, washed five times, and then incubated for 30 min at 37°C in RPMI medium containing 0.5% fatty acid–free BSA. Cells were seeded in the upper chambers of transwells (3421; Corning) and tested for transmigration across filters with 5-µm pores for 3 h at 37°C in response to the following chemoattractants in the lower chambers: CXCL12, CXCL13, CCL19, 7α,25-HC, or S1P (Sigma-Aldrich). To assess the role of MAPKs in B cell chemotaxis, cells were pretreated with ERK inhibitor U0126 (Merck Millipore) or p38 inhibitor SB203580 (Sigma-Aldrich) for 10 min and seeded in the transwells with the lower chambers containing CXCL12 (300 ng/ml) and the same concentrations of the inhibitor used for pretreatment. B cells that migrated to the lower chambers were enumerated by flow cytometry, and chemotactic responses were determined as percentages of their numbers relative to the numbers of input cells.
4
Compound Library Screening Protocol
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Thapsigargin (Tg), puromycin dihydrochloride, p38 inhibitor (SB203580), and JNK inhibitor (SP600125) were purchased from Sigma (St. Louis, MO, USA). AZD8055 and refametinib were procured from Cayman Chemical (Ann Arbor, MI, USA). The anti-puromycin and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies were purchased from Millipore (Burlington, MA, USA) and Santa Cruz Biotechnology (Dallas, TX, USA), respectively. The anti-phospho 4E-BP1 (Ser65), anti-4E-BP1, anti-phospho ERK 1/2, anti-ERK 1/2, anti-phospho p38, anti-p38, anti-phospho-JNK, anti-JNK, anti-phospho eIF2α, anti-eIF2α, anti-phospho c-Jun, anti-c-Jun, and horseradish peroxidase-conjugated secondary antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Alexa Fluor® 488 AffiniPure Goat Anti-Mouse IgG (Fcγ fragment specific) was supplied by Jackson ImmunoResearch Laboratories (West Grove, PA, USA). The Selleck Anti-Cancer Compound Library consisting of 414 drugs was purchased from the Department of Convergence Medicine, ASAN Medical Center, University of Ulsan College of Medicine (Seoul, Korea).
5
Modulation of Inflammatory Signaling
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Polyclonal anti-human IL-8 was obtained from R&D Systems (Minneapolis, MN, USA). IgG from goat serum was purchased from Sigma-Aldrich, Co. (St. Louis, MO, USA). Antibody to anti-p-JNK, anti-p-p38, anti-p38 and anti-IκB-α antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and anti-JNK, anti-p-IκB-α (ser32) was purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-tubulin antibody was purchased from Sigma-Aldrich, Co. Horseradish peroxidase (HRP)-conjugated secondary antibodies for immunoblotting were obtained from Santa Cruz Biotechnology. The NF-κB inhibitor BAY 11-7092, JNK inhibitor SP600125, p38 inhibitor SB203580 were purchased from Sigma-Aldrich, Co.
6
MAPK15 Regulation of c-Jun Phosphorylation
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AGS cells with low copy number were transfected with the pCMV6-Entry vector (0.8 μg/ml) that expresses Myc-DDK tagged MAPK15 protein or mock. On the 3rd day post-transfection, the cells were treated with 40μM JNK inhibitor SP600125 (Sigma) for 1 hour, 20μM MEK1/2 inhibitor U0126 (Sigma) for 2 hours, or 20μM p38 inhibitor SB203580(sigma) for 2 hours. After stimulation with culture medium including 20% FBS for 30 minutes, total protein was harvested and c-Jun and phosphorylated-c-Jun levels were detected by western blot analysis.
7
Antibody Reagents for Western Blot Analysis
8
Multiparameter Flow Cytometry Phenotyping
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mAbs specific for mouse B220 (RA3-6B2), c-Kit (2B8), CD3ε (145-2C11), CD5 (53-7.3), CD8 (53-6.7), CD11b (M1/70), CD11c (HL3), CD16/CD32 (2.4G2), CD19 (1D3), CD25 (PC61), CD34 (RAM34), CD45.1 (A20), CD45.2 (104), CD49b (DX5), CD127 (A7R34), GATA3 (L50-823), Gr-1 (RB6-8C5), MHC class II (M5/114.1), NK1.1 (PK136), Sca-1 (D7), Siglec-F (E50-2440), ST2 (U29-93), Thy1.2 (53-2.1), IL-4 (11B11), IL-12 (C15.6), IFN-γ (XMG1.2), and fluorochrome-conjugated streptavidin were purchased from BD Biosciences. mAbs specific for mouse CD4 (GK1.5), Flt3 (A2F10), F4/80 (BM8), IL-5 (TRFK5), IL-17RB (9B10), and NKp46 (29A1.4) were purchased from BioLegend. mAbs specific for mouse α4β7 (DATK32), FcεRIα (MAR-1), IL-13 (eBio13A), and killer cell lectin-like receptor G1 (KLRG1; 2F1) were purchased from eBioscience. mAbs against mouse CD16/CD32 (2.4G2), CD28 (37.51), and erythroid cell marker (TER-119) were purified from hybridoma culture supernatants in our laboratory.
Recombinant mouse IL-2, mIL-4, mIL-6, mIL-7, mIL-33, and mTGF-β1 were purchased from R&D Systems. p38 inhibitor (SB203580) and actinomycin D were purchased from Sigma-Aldrich.
Recombinant mouse IL-2, mIL-4, mIL-6, mIL-7, mIL-33, and mTGF-β1 were purchased from R&D Systems. p38 inhibitor (SB203580) and actinomycin D were purchased from Sigma-Aldrich.
9
Antibody Reagents for Western Blot Analysis
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Antibodies: anti-HA tag (clone 3F10) (Roche Applied Science), anti-β-actin and anti-alpha-tubulin (Sigma-Aldrich), anti-Smac (Upstate Cell Signaling Solutions, Lake Placid, NY), anti-Hsp60 (Enzo Life Sciences, Plymouth Meeting, PA), anti-VDAC1 (Calbiochem, Darmstadt, Germany), anti-GAPDH, anti-PDK4 and anti-Vinculin (Santa Cruz), anti-PGC-1α (a gift from Dr. Daniel P. Kelly, Sanford-Burnham Medical Research Institute, FL), anti-pAMPK, anti-pCREB and anti-CREB (Cell Signaling Technologies, Boston, MA), anti-CHCHD4 (Protein-Tech Group, IL), anti-Tim23 (BD Biosciences, San Diego, CA). The peroxidase-conjugated goat anti-rat, goat anti-rabbit, goat anti-mouse and horse anti-goat antibodies were from Vector Laboratories (Burlingame, CA). Rabbit polyclonal antibodies specific for human CHTM1 were generated in our laboratory through ProSci Inc. (Poway, CA) using full-length recombinant human CHTM1 protein purified from Escherichia coli. For cell transfections, Mirus (Madison, WI) and Lipofectamine 2000 (Invitrogen, Carlsbad, CA) were used. Restriction endonucleases were from New England BioLabs (Ipswich, MA). PKA inhibitor-H89, p38 inhibitor-SB203580, PI3K inhibitor-LY294002 and PKC inhibitor-GO6983 were from Sigma-Aldrich (St. Louis, MO). Other chemical reagents were from Thermo Fisher Scientific and Sigma-Aldrich.
10
MMP-1 and MMP-3 Regulation by EF Extract
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EF water extract was purchased from the Korean Plant Extract Bank (CW02-075; Daejeon, Korea), and a 50 mg/ml stock was prepared in distilled water. Human MMP-1 and MMP-3 antibodies and recombinant human IL-1β were obtained from R&D Systems (Minneapolis, MN, USA). Anti-β-actin antibody, p38 inhibitor SB203580, JNK inhibitor SP600125, and ERK inhibitor PD98059 were purchased from Sigma-Aldrich (St. Louis, MO, USA). p-c-Jun antibody was purchased from Abcam (Cambridge, MA, USA). PCNA, p65, and p50 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). p-STAT3, STAT-3, p-JNK, JNK, p-p38, p38, p-ERK, and ERK antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). High-glucose DMEM was purchased from HyClone (Logan, UT, USA). FBS and phosphate-buffered saline (PBS) were purchased from Gibco BRL (Gaithersburg, MD, USA).
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