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Phospho p38 mapk

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Phospho-p38 MAPK is a lab equipment product that detects the phosphorylated form of the p38 mitogen-activated protein kinase (MAPK). It is used to monitor the activation of the p38 MAPK signaling pathway.

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Lab products found in correlation

P38 mapk, by Cell Signaling Technology (118 mentions) Phospho jnk, by Cell Signaling Technology (37 mentions) Phospho akt, by Cell Signaling Technology (36 mentions) Phospho erk1 2, by Cell Signaling Technology (33 mentions) Erk1 2, by Cell Signaling Technology (27 mentions) β actin, by Merck Group (26 mentions) β actin, by Cell Signaling Technology (24 mentions) Phospho erk, by Cell Signaling Technology (23 mentions) Phospho sapk jnk, by Cell Signaling Technology (20 mentions) Gapdh, by Cell Signaling Technology (18 mentions) Cleaved caspase 3, by Cell Signaling Technology (16 mentions) Phospho p44 42 mapk erk1 2, by Cell Signaling Technology (15 mentions) Phospho nf κb p65, by Cell Signaling Technology (15 mentions) Caspase 3, by Cell Signaling Technology (14 mentions) Nf κb p65, by Cell Signaling Technology (13 mentions) Total p38 mapk, by Cell Signaling Technology (13 mentions) Gapdh, by Santa Cruz Biotechnology (11 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (11 mentions) Pvdf membrane, by Merck Group (11 mentions) Phospho p44 42 mapk, by Cell Signaling Technology (10 mentions)

Close spelling variants of this product

Phospho-p38 (477 citations) P38 MAPK (429 citations) Anti-phospho-p38 MAPK (111 citations) Phospho-p38 MAPK (Thr180/Tyr182) (64 citations) Anti-phospho-p38 MAPK (Thr180/Tyr182) (35 citations) Rabbit anti-phospho-p38 MAPK (19 citations) Phospho-p38 MAPK antibody (8 citations) Phospho-p38 MAPK (Thr180/Tyr182) Antibody (5 citations) Mouse anti-phospho-p38 MAPK (4 citations) Anti‐phospho‐p38 MAPK (T180/Y182) (4 citations)

248 protocols using phospho p38 mapk

1

Immunoblotting Muscle Protein Expression

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Immunoblots were performed as previously described.13 Homogenized tibialis anterior (TA) or soleus (20 μg per well) were separated and transferred to PVDF membranes (Millipore). Membranes were incubated with primary antibodies: phospho‐p38 MAPK (#9211, Thr180/Tyr182, Cell Signaling), p38 MAPK (#9212, Cell Signaling), RyR1 (#ab2868), CSQ1,2 (#ab3516), and DHPR (#ab2864) and thereafter with infrared‐labelled secondary antibodies (IRDye 680, IRDye 800, LI‐COR Biosciences). Detection and analyses were performed with the LI‐COR imaging system and normalized to total protein, which was determined by Ponceau S or Coomassie staining.
Mader T., Chaillou T., Alves E.S., Jude B., Cheng A.J., Kenne E., Mijwel S., Kurzejamska E., Vincent C.T., Rundqvist H, & Lanner J.T. (2022). Exercise reduces intramuscular stress and counteracts muscle weakness in mice with breast cancer. Journal of Cachexia, Sarcopenia and Muscle, 13(2), 1151-1163.
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2

Quantification of Phosphorylated Signaling Proteins

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Cells were harvested 24 hrs after seeding on PUR substrates in a radioimmunoprecipitation buffer containing a cocktail of protease and phosphatase inhibitors (Pierce). Equal protein concentrations were prepared for loading with NuPAGE sample buffer (Life Technologies) and separated on a 10% SDS-PAGE gel (BioRad). Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBS containing 0.1% Tween-20 for 1h at room temperature, followed by incubation with either phospho-p38MAPK (1:1000, Cell Signalling), p38MAPK (1:1000, Cell Signalling), phospho-Smad2/3 (1:1000, Cell Signalling) or Smad2/3 (1:1000, Cell Signalling) antibodies overnight at 4°C. After washing, membranes were blotted with anti-rabbit IgG (1:2000, SantaCruz), and bands were detected by enhanced chemiluminescence using an In-Vivo MS FX Pro (Bruker). Membranes were then stripped and reprobed using an antibody for β-actin (1:5000, Sigma) as a loading control. Phosphorylated events were quantified by normalizing the band intensity of phosphorylated protein to total protein. Analysis was performed using Image J software.
Page J.M., Merkel A.R., Ruppender N.S., Guo R., Dadwal U.C., Cannonier S., Basu S., Guelcher S.A, & Sterling J.A. (2015). Matrix Rigidity Regulates the Transition of Tumor Cells to a Bone-Destructive Phenotype through Integrin β3 and TGF-β Receptor Type II. Biomaterials, 64, 33-44.
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3

Signaling Pathway Activation in Macrophages

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WT and Dgkζ-/- BMMs treated with 100 ng/ml LPS, 100ng/ml IFNγ or 50 ng/ml IL-4 for the indicated time points were lysed in RIPA buffer (20 mM Tris-HCl, pH 7.5, 150 mM NaCl 1 mM EDTA, 1 mM EGTA, 1% Nonidet P-40, 1% sodium deoxycholate) supplemented with protease/phosphatase inhibitor cocktail (Pierce). The protein lysates were resolved by SDS-PAGE and electro-transferred into nitrocellulose membranes and probed with specific antibodies. The following antibodies were used: phospho-STAT1 (7649, Cell Signaling Technology), STAT1 (9172, Cell Signaling Technology), phospho-STAT3 (9145, Cell Signaling Technology), STAT3 (4904, Cell Signaling Technology), phospho-STAT6 (9361, Cell Signaling Technology), STAT6 (5397, Cell Signaling Technology), phospho-AKT (4060, Cell Signaling Technology), AKT (2966, Cell Signaling Technology), phospho-p38 MAPK (9216, Cell Signaling Technology), p38 MAPK (9219, Cell Signaling Technology), phospho-GSK3β (9336, Cell Signaling Technology), GSK3β (9315, Cell Signaling Technology) and β-Actin (A5441, Sigma).
Mahajan S., Mellins E.D, & Faccio R. (2019). Diacylglycerol kinase ζ regulates macrophage responses in juvenile arthritis and cytokine storm syndrome mouse models. Journal of immunology (Baltimore, Md. : 1950), 204(1), 137-146.
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4

Western Blot Analysis of Protein Expression

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Cells pellets were lysed in RIPA buffer (Sigma-Aldrich). Supernatant containing 20 µg of proteins was resolved in SDS-polyacrylamide gel and electroblotted onto PVDF membranes (Millipore, Billerica, MA) using a semidry transfer unit (ATTO, Tokyo, Japan). Immunoblotting was performed with anti-VEGF, MMP-2, Vimentin (Santa Cruz), N-Cadherin, uPA & hnRNP-K (Abcam), phospho-p38MAPK, E-Cadherin (Cell Signaling), HSP27 (StressMarq Biosciences Inc.) and β-actin (Abcam, Cambridge) antibodies. The membranes probed with the first antibodies were excessively washed with TBS-T (Tris-buffered saline-Tween 20) and incubated with secondary horseradish peroxidase (HRP)-conjugated goat anti-mouse or anti-rabbit (Santa Cruz) antibodies. Protein bands were detected using ECL prime substrate (GE Healthcare, CA). Densitometric quantitation of three independent immunoblotting experiments was performed with the Image J software (NIH, Bethesda, MD). Expression level of each of the proteins in control and treated cells was calculated with respect to the β-actin (loading control). All experiments were performed in triplicate.
Chaudhary A., Kalra R.S., Malik V., Katiyar S.P., Sundar D., Kaul S.C, & Wadhwa R. (2019). 2, 3-Dihydro-3β-methoxy Withaferin-A Lacks Anti-Metastasis Potency: Bioinformatics and Experimental Evidences. Scientific Reports, 9, 17344.
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5

Comprehensive Signaling Pathway Analysis

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PTEN(9559), p44/42(Erk1/2)(9107), phospho-p44/42(Thr202/Tyr204)(9101), p38MAPK(8690), phospho-p38MAPK(Thr180/Tyr182)(4511), eIF4E(9742), phospho-eIF4E(Ser209)(9741), Mnk(2195), phospho-Mnk(Thr197/202)(2111), phospho-4EBP1(Ser65)(9451), AKT(4685), phospho-Akt(Ser473)(9271), mTOR(2983), phospho-mTOR(Ser2448)(2971), phospho-p70S6K(Thr421/Ser424)(9205), AR(3202), α-Tubulin(2125), c-Myc(9402), Survivin(2808), Cyclin D1(2978), BCL-2(2872), p-eIF4G(2441) and EGFR(2232) were from Cell Signaling Technology (Beverly, MA), 4EBP1(6936), AR(N-20)(816), β-Actin(130656) were from Santa Cruz Biotechnology (Santa Cruz, CA). GAPDH(MAB374) was from EMD Millipore, Darmstadt, Germany. phospho-eIF4E(Ser209)(ab76256) for immunohistochemistry and immunofluorescence were from Abcam, Cambridge, UK. Ki67(7240) antibody was from DAKO via Agilent Technologies, Santa Clara, CA.
D’Abronzo L.S., Bose S., Crapuchettes M.E., Beggs R.E., Vinall R.L., Tepper C.G., Siddiqui S., Mudryj M., Melgoza F.U., Durbin-Johnson B.P., deVere White R.W, & Ghosh. P.M. (2017). The Androgen Receptor is a negative regulator of eIF4E Phosphorylation at S209: Implications for the use of mTOR inhibitors in advanced prostate cancer. Oncogene, 36(46), 6359-6373.
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6

Protein Extraction and Immunoblotting for RBL-2H3 Cells

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Nuclear and cytosolic proteins were extracted as previously described19 (link). Before protein extraction, RBL-2H3 cells (2 × 106/well in 6-well plates) were sensitized with anti-DNP IgE (50 ng/ml). After incubation overnight, the cells were pretreated with or without PD for 1 h and challenged with DNP-HSA (100 ng/ml). Then, cell extracts were prepared by detergent lysis procedure as described previously19 (link). Samples of protein (30 μg) were electrophoresed using 8–12% SDS-PAGE gel at 120 V for 90 min and transferred to polyvinylidene difluoride membranes (Amersham Pharmacia Biotech, Piscataway, NJ, USA). Immunodetection was carried out using a chemiluminescent substrate. The antibodies of IKK, phospho-IKK, NF-κB p65, phospho-NF-κB p65, PARP, IκBα, phospho-IκBα, and β-actin were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The antibodies of Syk, phospho-Syk, Lyn, phospho-Lyn, Gab2, phospho-Gab2, Akt, phospho-Akt, p38 MAPK, phospho-p38 MAPK, ERK, phospho-ERK, Nrf-2, and HO-1 were purchased from Cell Signaling Technology (Danvers, MA, USA).
Ye J., Piao H., Jiang J., Jin G., Zheng M., Yang J., Jin X., Sun T., Choi Y.H., Li L, & Yan G. (2017). Polydatin inhibits mast cell-mediated allergic inflammation by targeting PI3K/Akt, MAPK, NF-κB and Nrf2/HO-1 pathways. Scientific Reports, 7, 11895.
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7

Western Blot Analysis of Signaling Proteins

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Western blotting was performed following standard protocols. Primary antibodies reactive to phospho-NF-kB p65 (1:1000; Cell Signaling, 3033), NF-kB p65 (1:1000; Cell Signaling, 4764), phospho-AKT (1:500; Cell Signaling, 4060), AKT (1:1000; Cell Signaling, 4685), phospho-p38 MAPK (1:800; Cell Signaling, 4631), p38 MAPK (1:1000; Cell Signaling, 9212), PGC1β (1:1000; Abcam, 176328), and β-actin (1:20,000; Sigma, F3022) were used. Primary antibody binding was detected by enhanced chemoluminescence (GE Healthcare, RPN2106).
Rao S., Sigl V., Wimmer R.A., Novatchkova M., Jais A., Wagner G., Handschuh S., Uribesalgo I., Hagelkruys A., Kozieradzki I., Tortola L., Nitsch R., Cronin S.J., Orthofer M., Branstetter D., Canon J., Rossi J., D'Arcangelo M., Botling J., Micke P., Fleur L.L., Edlund K., Bergqvist M., Ekman S., Lendl T., Popper H., Takayanagi H., Kenner L., Hirsch F.R., Dougall W, & Penninger J.M. (2017). RANK rewires energy homeostasis in lung cancer cells and drives primary lung cancer. Genes & Development, 31(20), 2099-2112.
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8

Analyzing Protein Expression Levels

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To analyze protein expression levels, cells were washed briefly in phosphate buffered saline (PBS), lysed in urea sample buffer (8 M deionized urea, 1% SDS, 10% Glycerol, 60 mM Tris pH 6.8, and 5% beta-mercaptoethanol) and equalized for total protein concentration. Samples were subjected to SDS-PAGE (Genscript, Piscataway, NJ), followed by transfer to PVDF membranes (Sigma). Membranes were probed with the following antibodies: Desmoplakin / NW6, Plakoglobin / 1407 (Kathleen Green, Northwestern University), GAPDH, Plakophilin 2, RhoA (Santacruz Biotechnology, Dallas, TX), p38 MAPK, phospho-p38 MAPK (Cell Signaling Technology, Danvers, MA), Rac1 (BD Biosciences, San Jose, CA), Desmoglein-2 / 6D8 (ThermoFisher) and HRP-conjugated secondary antibodies from Jackson ImmunoResearch (West Grove, PA). Blots were visualized by enhanced chemiluminescence using a ChemiDoc imager (BioRad, Hercules, CA). All western blots shown are representative of data from three independent experiments. To measure levels of GTP-bound RhoA and Rac1, multiple scratch wounds were made in triplicate samples of control and DP knockdown cells, and colorimetric G-LISA assays performed according to manufacturer’s instructions (Cytoskeleton, Denver CO). Absorbance measurements at 490 nm (indicative of GTPase activity) were obtained using an Epoch 2 plate reader (Biotek, Winooski, VT).
Bendrick J.L., Eldredge L.A., Williams E.I., Haight N.B, & Dubash A.D. (2018). Desmoplakin harnesses Rho GTPase and p38 MAPK signaling to coordinate cellular migration.. The Journal of investigative dermatology, 139(6), 1227-1236.
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9

Western Blot Analysis of EMT Markers

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Equal quantities of cell extracts were separated on a 10% SDS-PAGE gel, transferred, and analyzed by the Western lightning plus-ECL detection system (Perkin Elmer, Inc., Waltham, MA, USA). Briefly, antibodies against NDRG1, NDRG2, NDRG3 (Invitrogen), Snail (Abcam, Cambridge, UK), cyclin E, E-cadherin, STAT3, N-cadherin, Vimentin (Abgent, San Diego, CA, USA), Slug, p44/42 MAPK, Phospho-p44/42 MAPK, SAPK/JNK, Phospho-SAPK/JNK, p38 MAPK, Phospho-p38 MAPK (Cell Signaling, Danvers, MA, USA), Phospho-STAT3 and β-actin (Millipore) were used and the protein bands were detected and quantified with the ChemiGenius Image Capture System (Syngene, Cambridge, UK) and the GeneTools Program of ChemiGenius (Syngene).
Chiang K.C., Yang S.W., Chang K.P., Feng T.H., Chang K.S., Tsui K.H., Shin Y.S., Chen C.C., Chao M, & Juang H.H. (2018). Caffeic Acid Phenethyl Ester Induces N-myc Downstream Regulated Gene 1 to Inhibit Cell Proliferation and Invasion of Human Nasopharyngeal Cancer Cells. International Journal of Molecular Sciences, 19(5), 1397.
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10

Dissecting Apoptosis Signaling Pathways

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G1 was purchased from Tocris (Wiesbaden-Nordenstadt, Germany), dissolved (5 mM) in dimethyl sulfoxide (DMSO) (Roth, Karsruhe, Germany) and stored at −20 °C; Thapsigargin, SP600125, GSK2606414 were also purchased from Tocris. Indo-1 AM was from Thermo Fisher Scientific (Waltham, MA, USA). zVAD-fmk was bought from Santa Cruz (Santa Cruz, CA, USA). SB203580 and Kira6 were purchased from MERCK Millipore (Darmstadt, Germany). All substances were dissolved in DMSO. Antibodies were obtained from the following commercial sources: caspase 9 (Ca# 9502), cleaved PARP (Ca# 9541), IRE1α (Cat# 3294), PERK (Cat# 3192), eIF2α (Cat# 5234), phospho-eIF2α (Cat# 3398), BiP GRP78 (Ca# 3177), CHOP (Cat# 2895), p38 MAPK (Ca# 9212), phospho-p38 MAPK (Ca# 4511), phospho-SAPK/JNK (Ca# 4668), caspase 3 (Ca# 9662), BCL-2 (Ca# 2872), Cell Signaling (Danvers, MA, USA); ATF6 (Cat# 73500), BioAcademia (Osaka, Japan); puromycin (Ca# MABE343), cylophilin D (Ca# AP1035), MERCK Millipore (Darmstadt, Germany); phospho-IRE1α (Cat# NBP2-50067), Novus Biologicals (Littleton, CO, USA); cytochrome c (Ca# 556433), BD Biosciences (Franklin Lakes, NJ, USA); β-actin (Cat# A5441, Sigma-Aldrich (Steinheim, Germany)). Secondary, peroxidase-conjugated antibodies were purchased from Dianova (Hamburg, Germany). All other chemicals of analytical grade were obtained from Sigma-Aldrich or Roth.
Vo D.K., Hartig R., Weinert S., Haybaeck J, & Nass N. (2019). G-Protein-Coupled Estrogen Receptor (GPER)-Specific Agonist G1 Induces ER Stress Leading to Cell Death in MCF-7 Cells. Biomolecules, 9(9), 503.
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