Phospho p38

Manufactured by Santa Cruz Biotechnology

Sourced in United States, United Kingdom

Phospho-p38 is an antibody product designed to detect the phosphorylated form of the p38 mitogen-activated protein kinase (MAPK) protein. Phosphorylation of p38 MAPK is a key event in the activation of this important signaling pathway. The Phospho-p38 antibody can be used to monitor the activation state of the p38 MAPK in various cellular and experimental contexts.

Automatically generated - may contain errors

Lab products found in correlation

40 protocols using phospho p38

TLR Agonist-Mediated Immune Responses

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

S. aureus (strain RN6390) [11 (link),12 (link),13 (link),14 (link)] was maintained in tryptic soy broth (TSB; Sigma-Aldrich, St. Louis, MO). Bacterial lipopeptide Pam3Cys-Ser-(Lys)4 hydrochloride (Pam3Cys, TLR1/2 agonist), polyI:C (TLR3 agonist), Lipoploysacchride (LPS, TLR4 agonist), flagellin (TLR5 agonist), PolydT (TLR7/8 agonist), and ODN (TLR9 agonist) were purchased from InvivoGen (San Diego, CA). Antibodies against p-ERK, ERK, phospho-p38, p38, IkB-α, and TLR 3, 4, 5, 7, and 9 were purchased from Santa Cruz Biotechnology Inc. (CA, USA). Anti-phospho-IkB-α and anti-TLR2 antibodies were purchased from Cell Signaling Technology (Beverly, MA). A mouse monoclonal anti-β-actin antibody was purchased from Sigma (St. Louis, MO). Secondary horseradish peroxidase (HRP)-conjugated anti-mouse or anti-rabbit IgG antibodies were purchased from Bio-Rad (Hercules, CA).

Singh P.K, & Kumar A. (2015). Retinal Photoreceptor Expresses Toll-Like Receptors (TLRs) and Elicits Innate Responses Following TLR Ligand and Bacterial Challenge. PLoS ONE, 10(3), e0119541.

+ Open protocol

+ Expand

Western Blot Analysis of T Cell Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

A total of 5×10⁶ CD4⁺ T cells in each group were extracted from the cytosolic and nuclear fractions using cell lysis buffer. The supernatants were separated, and the protein concentration in the fractions was quantified using a Bradford quantitative protein assay kit (Applygen Technologies Inc., China). The total protein lysates (40 μg per each well) were loaded and subjected to 10-12% SDS-PAGE before being electro-transferred to polyvinylidene difluoride membranes (PVDF) at 100 V for 1 h. The membrane was blocked with 5% nonfat milk in Tris-buffered saline supplemented with 0.05% Tween 20 (pH 7.6) at 4°C for 2 h and incubated with anti-mouse and anti-human polyclonal antibodies, including antibodies against ERK, phospho-ERK, JNK, phospho-JNK, NFAT1, p65 (NF-κB), phospho-p65 (NF-κB), p38, phospho-p38 and β-actin (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA and Cell Signaling, USA), overnight at 4°C. The membrane was incubated with horseradish peroxidase-conjugated anti-mouse and anti-rabbit secondary antibody (1:2,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA) at room temperature for 2 h. After extensive washing, the blot was developed using an ECL chemiluminescent detection kit (TransGen Biotech Co. Ltd., China) according to the manufacturer's instructions. The X-ray films were scanned and quantified by Gel-Doc (Bio-red) using image analysis software.

Samivel R., Kim D.W., Son H.R., Rhee Y.H., Kim E.H., Kim J.H., Bae J.S., Chung Y.J., Chung P.S., Raz E, & Mo J.H. (2015). The role of TRPV1 in the CD4+ T cell-mediated inflammatory response of allergic rhinitis. Oncotarget, 7(1), 148-160.

+ Open protocol

+ Expand

Quercetin Modulation of Cell Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Dulbecco’s modified Eagle’s media (DMEM), Dulbecco’s phosphate buffer saline (DPBS), fetal bovine serum (FBS), penicillin–streptomycin, quercetin dihydrate (Qu), 3-(4,5-dimetylthiazol-yl)-diphenyl tetrazolium bromide (MTT), propidium iodide, rhodamine 123, 2,7-dichlorodihydrofluoresceindiacetate (H₂DCF-DA), Fluo-3 AM, proteinase K, ribonuclease A, 1,2-bis-(O-aminophenoxy)-ethane-N,N,N’,N’-tetraacetic acid, tetraacetoxymethyl ester (BAPTA-AM), β-nicotinamide adenine dinucleotide reduced disodium salt (β-NADH), ascorbic acid, agarose, dimethyl sulfoxide (DMSO) and anti-β-actin antibody were purchased from Sigma–Aldrich Chemicals (St. Louis, MO). Antibodies against Bcl-2, Bax, Bim, AIF, NF-κB, caspase-3/caspase-8, phospho-MEK, MEK1/2, phospho-ERK, ERK 1/2, phospho-p38, p-38, phospho-JNK, JNK, Nrf-2, catalase, Cu/Zn SOD, PI3K, phospho-Akt, Akt and GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Inc,). FITC Annexin V was purchased from BD Pharmingen. Histone H3 antibody was purchased from Cell Signaling Technology.
Stock solution of quercetin (Qu) was prepared in dimethyl sulfoxide (DMSO) and diluted to the desired final concentration with culture medium just before use. The final DMSO concentration did not exceed 0.1% (v/v).

Rafiq R.A., Quadri A., Nazir L.A., Peerzada K., Ganai B.A, & Tasduq S.A. (2015). A Potent Inhibitor of Phosphoinositide 3-Kinase (PI3K) and Mitogen Activated Protein (MAP) Kinase Signalling, Quercetin (3, 3', 4', 5, 7-Pentahydroxyflavone) Promotes Cell Death in Ultraviolet (UV)-B-Irradiated B16F10 Melanoma Cells. PLoS ONE, 10(7), e0131253.

+ Open protocol

+ Expand

Cinobufagin-Induced Apoptosis and Autophagy

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cinobufagin was obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China) and dissolved in Dulbecco's modified Eagle's medium (DMEM) (Gibco, Grand Island, NY, USA) to each working dose. Fetal bovine serum (FBS) was purchased from Hangzhou Sijiqing Biological Engineering Material Co., Ltd. (Hangzhou, Zhejiang, China). Chloroquine (CQ), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), N-acetyl cysteine (NAC) and 3-methyladenine (3-MA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The Annexin V-FITC Apoptosis Detection Kit I was obtained from BD Biosciences (San Diego, CA, USA). Hoechst 33258 was purchased from Promega Corp. (Madison, Wi, USA). Lipofectamine 2000 transfection reagent was purchased from Invitrogen Life Technologies (Carlsbad, CA, USA). Antibodies against caspase-3, caspase-8, caspase-9, cleaved PARP, phospho-JNK, total JNK, phospho-p38, total p38, LC3B, p62, GAPDH, enhanced chemiluminescence (ECL) and western blotting kits were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

MA K., ZHANG C., HUANG M.Y., LI W.Y, & HU G.Q. (2016). Cinobufagin induces autophagy-mediated cell death in human osteosarcoma U2OS cells through the ROS/JNK/p38 signaling pathway. Oncology Reports, 36(1), 90-98.

+ Open protocol

+ Expand

Protein Expression Profiling in Heart Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols

Heart tissues were removed from liquid nitrogen, thawed, and weighed. Tissue homogenate was prepared with RIPA buffer (150 mM NaCl, 10% Triton X-100, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulphate, and 50 mM Tris base), supplemented with protease inhibitor cocktail (Sigma Aldrich, USA). Then homogenate was centrifuged at 12000 rpm for 15 min at 4°C. Protein concentration was measured by the method described by Bradford [20 (link)]. Each well was loaded with equal amount of proteins (40 μg) in a sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to nitrocellulose membrane. Following this, membrane was probed with primary antibodies, that is, ERK1/ERK2 (Cell signaling, USA), phospho-ERK1/ERK2 (Thr202/Tyr204; Cell signaling, USA), SAPK/JNK (Cell signaling, USA), phospho SAPK/JNK (Thr183/Tyr185; Cell signaling, USA), p38 (Abcam; UK); phospho-p38 (Santa Cruz, USA), NFκBp65 (Cell signaling, USA), and β-actin (Cell signaling, USA) overnight at 4°C. These primary antibodies were detected by adding HRP-conjugated secondary antibodies (Merck Genei, India) after incubating for 2 h at room temperature. Later, the bound antibodies were visualized with an enhanced chemiluminescence (ECL) (Thermo Fischer Scientific Inc., USA) kit and quantified by densitometric analysis.

Suchal K., Malik S., Gamad N., Malhotra R.K., Goyal S.N., Chaudhary U., Bhatia J., Ojha S, & Arya D.S. (2016). Kaempferol Attenuates Myocardial Ischemic Injury via Inhibition of MAPK Signaling Pathway in Experimental Model of Myocardial Ischemia-Reperfusion Injury. Oxidative Medicine and Cellular Longevity, 2016, 7580731.

+ Open protocol

+ Expand

Protein Expression Analysis in Kidney Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary kidney cells and human podocytes were lysed in RIPA buffer and protein estimation was performed using the BCA method. 20μg protein samples from lysates were used for western blotting analysis. Myo1c, Neph1, Nephrin and Myo1b antibodies have been described previously.^{18 (link),31 (link),32 (link)} Other antibodies, including GAPDH (Sigma # G8795), α-SMA (Santa Cruz # c-53142), NM1 (Sigma # M3567), phospho-SMAD2 (Invitrogen # 44–244G), phspho-SMAD3 (Millipore # 07–1389), SMAD2/3 (Cell Signaling # 3102S), phospho-p38 (Santa Cruz # sc-166182), p38 MAPK (Cell Signaling # 9212S), phospho-JNK (Cell Signaling # 9255S), JNK (Cell Signaling # 9252S), phospho-ERK1/2 (Cell Signaling # 9101), ERK1/2 (Cell Signaling # 4695S), GDF-15 Antibody (Santa Cruz # sc-377195) and JunB Antibody (Santa Cruz # sc-8051) were commercially obtained. Western blotting image acquisition and densitometric analysis was performed using LI-CORE imaging station. The fluorophore secondary antibodies, Alexa Fluor 488 goat anti-rabbit IgG (H+L), Alexa Fluor 568 goat anti-mouse IgG (H+L) and Alexa Fluor phalloidin were commercially obtained from Invitrogen. Doxorubicin hydrochloride (adriamycin) was purchased from R&D Systems, Inc. Nephrotoxic serum was obtained from Dr. David Salant^{74 (link)} and Probetex Inc. (PTX-001) and used as previously described.^{26 (link),36 (link)}

Arif E., Solanki A.K., Srivastava P., Rahman B., Tash B.R., Holzman L.B., Janech M.G., Martin R., Knölken H.J., Fitzgibbon W.R., Deng P., Budisavljevic M.N., Syn W.K., Wang C., Lipschutz J.H., Kwon S.H, & Nihalani D. (2019). The motor protein Myo1c regulates transforming growth factor-β-signaling and fibrosis in podocytes. Kidney international, 96(1), 139-158.

+ Open protocol

+ Expand

Curcumin-Lecithin Antioxidant Activity Study

Check if the same lab product or an alternative is used in the 5 most similar protocols

Curcuma Longa Linn (powdered form) and lecithin (L-α-phosphatidylcholine) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The organic solvents such as toluene and dichloromethane were purchased from Fisher Scientific (Waltham, MA, USA). Fetal bovine serum (FBS) and phosphate-buffered saline (PBS) were purchased from GE Healthcare (Logan, UT, USA). The following antibodies were obtained: c-Src, phospho-c-Src, PKC, phospho-PKC, JNK, phospho-JNK, p38, phospho-p38, ERK, phospho-ERK, IκBα, phospho-IκBα, NF-κBp65, phospho-NF-κBp65, Bcl-2, Bax, cleaved caspase-3, and β-actin antibodies (Santa Cruz Biotechnology, Paso Robles, CA, USA); The following reagents were obtained: N-acetylcysteine (NAC) (Tocris, KOMA Biotech, Seoul, Korea) and 5-(and-6)-chloromethyl-2′,7′- dichlorodihydrofluorescein diacetate, acetyl ester (CM-H₂DCFDA) (Invitrogen, Carlsbad, CA, USA). PP2, SP600125, Bisindolylmaleimide I, and Bay11-7082 were obtained from MedChemExpress (Monmouth Junction, NJ, USA). All other reagents did not show any critical cytotoxic effects by themselves.

Kim J.Y., Lee Y.M., Kim D.W., Min T, & Lee S.J. (2020). Nanosphere Loaded With Curcumin Inhibits the Gastrointestinal Cell Death Signaling Pathway Induced by the Foodborne Pathogen Vibrio vulnificus. Cells, 9(3), 631.

+ Open protocol

+ Expand

Protein Analysis of Brain Samples After SAH

Check if the same lab product or an alternative is used in the 5 most similar protocols

The brain samples were collected at 24 hours after SAH. Proteins of the ipsilateral cortex and hippocampus were extracted by homogenizing in RIPA buffer (Santa Cruz Biotechnology, USA). Western blotting was performed as described previously^{19 (link)} using FPR2 (Santa Cruz Biotechnology, USA), phospho-p38 (Santa Cruz Biotechnology), p38 (Santa Cruz Biotechnology), IL-1β (Abcam, USA) and IL-6 (Santa Cruz Biotechnology, USA).

Guo Z., Hu Q., Xu L., Guo Z.N., Ou Y., He Y., Yin C., Sun X., Tang J, & Zhang J.H. (2016). Lipoxin A4 reduces inflammation through formyl peptide receptor 2 /p38MAPK signaling pathway in subarachnoid hemorrhage rats. Stroke; a journal of cerebral circulation, 47(2), 490-497.

+ Open protocol

+ Expand

Western Blot Analysis of Cell Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysates were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the separated proteins were transferred onto pure nitrocellulose blotting membrane (Pall Gelman Laboratory, Ann Arbor, MI, USA). The membranes were incubated with primary antibodies against phospho-H2A.X (Ser139), CHOP, beclin-1, LC3B, caspase-3, caspase-9, ERK, JNK (Cell Signaling Technology, Beverly, MA, USA), GRP78, Bax, phospho-p38 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), phospho-IRE1 (Abcam, Cambridge, MA, USA), and actin (Sigma-Aldrich), followed by incubating with a corresponding secondary antibody (Pierce, Rockford, IL, USA). The Amersham enhanced chemiluminescence plus western blotting detection system (GE Healthcare Life Sciences, Buckinghamshire, UK) was used to detect the protein bands.

Zhen A.X., Piao M.J., Hyun Y.J., Kang K.A., Fernando P.D., Cho S.J., Ahn M.J, & Hyun J.W. (2019). Diphlorethohydroxycarmalol Attenuates Fine Particulate Matter-Induced Subcellular Skin Dysfunction. Marine Drugs, 17(2), 95.

+ Open protocol

+ Expand

Antibody Panel for Signaling Study

Check if the same lab product or an alternative is used in the 5 most similar protocols

Following antibodies were used in this study: BAD (H-168), phospho-AKT (S473), phospho-ERK1, ERK1, ERβ, Actin, Flag, phospho-c-Jun, c-Jun, phospho-JNK, JNK, phospho-p38, CXCR4, β-catenin, GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA); phospho-BAD (S136), Sp1 (Upstate, Lake Placid, NY); AKT (Cell Signaling, Danvers, MA); cyclin D1 (Novocastra Laboratories Ltd., Newcastle Upon Tyne, UK); SNAIL (Abcam, Cambridge, MA); GFP (Sigma, St. Louis, MO) and MTA3 antibody was described in [41 (link)]. Secondary antibodies: Amersham (San Diego, California) and Chemicon (Millipore, Billerica, MA).

Cekanova M., Fernando R.I., Siriwardhana N., Sukhthankar M., de la Parra C., Woraratphoka J., Malone C., Ström A., Baek S.J., Wade P.A., Saxton A.M., Donnell R.M., Pestell R.G., Dharmawardhane S, & Wimalasena J. (2014). BCL-2 family protein, BAD is down-regulated in breast cancer and inhibits cancer cell invasion. Experimental cell research, 331(1), 1-10.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!