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P p44 42 mapk erk1 2

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P-p44/42 MAPK (Erk1/2) is a laboratory reagent that detects the phosphorylated forms of the extracellular signal-regulated kinase 1 and 2 (ERK1/2) proteins. ERK1/2 are members of the mitogen-activated protein kinase (MAPK) family and play a key role in cellular signaling pathways.

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Lab products found in correlation

β actin, by Cell Signaling Technology (4 mentions) P44 42 mapk erk1 2, by Cell Signaling Technology (3 mentions) Cleaved caspase 3, by Cell Signaling Technology (3 mentions) Erk1 2, by Cell Signaling Technology (3 mentions) Pfrs2α, by Cell Signaling Technology (2 mentions) Caspase 3, by Cell Signaling Technology (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) P gsk3β ser9, by Cell Signaling Technology (1 mentions) P p38 thr180 tyr182, by Cell Signaling Technology (1 mentions) β actin, by Bioworld Technology (1 mentions) Phsl ser660, by Cell Signaling Technology (1 mentions) Ba1054, by Boster Bio (1 mentions) Gsk3β, by Cell Signaling Technology (1 mentions) Chemidoc imaging system, by Bio-Rad (1 mentions) Bca kit, by Keygen Biotech (1 mentions) Ab170099, by Abcam (1 mentions) P s6 ribosomal protein, by Cell Signaling Technology (1 mentions) Pathscan multiplex western cocktail 1, by Cell Signaling Technology (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Fgfr1, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

ERK1/2 (1487 citations) P-ERK1/2 (1094 citations) P44/42 MAPK (Erk1/2) (135 citations) P44/42 MAPK (90 citations) P-MAPK (38 citations) P-p44/42 MAPK (20 citations) Rabbit anti-phospho p44/42 MAPK (p- Erk1/2), (6 citations) ERK1/2 MAPK (3 citations) Anti-p-p44/42 MAPK (Erk1/2), (3 citations) ERK1/2 p44/42 (2 citations)

12 protocols using p p44 42 mapk erk1 2

1

Adipose Tissue Protein Analysis

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Epididymal adipose tissue was homogenized with a tissue grinder at 4°C temperature and the lysate were centrifugation at 12000 rpm for 20 minutes. The protein concentrations were measured with BCA kit (KeyGEN BioTECH Corp., Ltd, Nanjin, China). The protein samples were denatured, separated by SDS-PAGE and then transferred to the PVDF membrane. Membranes were blocked with 5% skim milk or bovine serum albumin (BSA) for 3 hours, and then incubated overnight at 4°C with specific primary antibodies as follows: ATGL (2138S, Cell Signaling Technology), P-HSL (ser660) (4126S, Cell Signaling Technology), HSL (4107S, Cell Signaling Technology), P-GSK-3β (ser9) (5558S, Cell Signaling Technology), GSK-3β (9315S, Cell Signaling Technology), β-actin (AP0060, Bioworld), PKA (5842S, Cell Signaling Technology), P-p38 (Thr180/tyr182) (9219S, Cell Signaling Technology), p38 (ab170099, Abcam), P44/42 MAPK (ERK1/2) (9102S, Cell Signaling Technology), P-P44/42 MAPK (ERK1/2) (Thr202/Tyr204) (Cell Signaling Technology) and AT2R (ab92445, Abcam). Membranes were washed and incubated with secondary antibodies (BA1054, BOSTER Biological Technology) at room temperature for 3 hours. After washing, membranes were exposed by Super ECL Prime (SEVEN BIOTECH) with ChemiDoc™ Imaging System (BIO-RAD, USA). The images were analyzed quantitatively by densitometry with Image J software.
Li A., Shi W., Wang J., Wang X., Zhang Y., Lei Z, & Jiao X.Y. (2022). The gene knockout of angiotensin II type 1a receptor improves high-fat diet-induced obesity in rat via promoting adipose lipolysis. PLoS ONE, 17(7), e0267331.
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2

Co-culture Assay for Prostate Cancer

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MDA PCa 118b cells were co-cultured with PMOs in a two-compartment system as previously reported (14 (link)). After 48 h of co-culture, PMO and PCa cell numbers were estimated by [3H]-thymidine incorporation assay (13 (link)). Western blotting was done by standard procedures with antibodies against p-FRS2α (Cell Signaling, rabbit polyclonal [Tyr196]) or with PathScan Multiplex Western Cocktail I against p-p90RSK, p-Akt, p-p44/42 MAPK (Erk1/2), and p-S6 ribosomal protein (Cell Signaling). For western blot analysis, dissected tumor bearing bones or bone tissues were frozen in liquid nitrogen and then pulverized with stainless-steel mortar and pestle. Tissue powder was then suspended in lysis buffer containing Protease Inhibitor Cocktail (Roche Applied Science) and, if phosphorylated proteins will be investigated, also Phosphatase inhibitor Cocktail (PhosSTOP EASYpack Roche). The suspension was vortexed and then homogenized by sonication. Supernatant fraction of lysates was then used for western blot analysis. Band densities on western blots were assessed with Image J software (National Institutes of Health).
Wan X., Corn P.G., Yang J., Palanisamy N., Starbuck M.W., Efstathiou E., Li-Ning Tapia E.M., Zurita A.J., Aparicio A., Ravoori M.K., Vazquez E.S., Robinson D.R., Wu Y.M., Cao X., Iyer M.K., McKeehan W., Kundra V., Wang F., Troncoso P., Chinnaiyan A.M., Logothetis C.J, & Navone N.M. (2014). Prostate Cancer Cell–Stromal Cell Cross-Talk via FGFR1 Mediates Antitumor Activity of Dovitinib in Bone Metastases. Science translational medicine, 6(252), 252ra122.
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3

Immunohistochemical Analysis of Subcutaneous PCa PDX Models

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For subcutaneous PCa PDXs, tumor samples were prepared as previously described (13 (link)). IHC analysis was done at described elsewhere (29 (link)), using antibodies against FGFR1 (Epitomics, rabbit monoclonal clone ID EPR806Y), FGFR1 (Cell Signaling, rabbit monoclonal), FRS2α (Santa Cruz Biotechnology, rabbit polyclonal sc83138), p-FRS2α (Cell Signaling, rabbit polyclonal (Tyr196)), p-p44/42 MAPK (Erk1/2) (Cell Signaling, rabbit monoclonal (Thr202/Tyr204)), p-AKT (Dako, rabbit monoclonal), p-S6K (p-S434) (AbCam, ab47379), androgen receptor (Dako, rabbit polyclonal) and cleaved caspase 3 (Cell Signaling, rabbit polyclonal [Asp175]).
Wan X., Corn P.G., Yang J., Palanisamy N., Starbuck M.W., Efstathiou E., Li-Ning Tapia E.M., Zurita A.J., Aparicio A., Ravoori M.K., Vazquez E.S., Robinson D.R., Wu Y.M., Cao X., Iyer M.K., McKeehan W., Kundra V., Wang F., Troncoso P., Chinnaiyan A.M., Logothetis C.J, & Navone N.M. (2014). Prostate Cancer Cell–Stromal Cell Cross-Talk via FGFR1 Mediates Antitumor Activity of Dovitinib in Bone Metastases. Science translational medicine, 6(252), 252ra122.
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4

Inhibition of Oncogenic Pathways

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I-BET726 was purchased from Adooq (Shanghai, China). MTT, AZD5153, PD98059 and LY294002, JQ1 and CPI203 were purchased from Sigma (Shanghai, China). Z-DEVD-fmk, Z-LEHD-fmk, and Z-VAD-fmk were provided by Calbiochem (La Jolla, CA). Antibodies for c-Myc (#9402), Cyclin D1 (#2922), BRD4 (#13440), Bcl-2 (#15707), SphK1 (#12071), phosphorylated (“p”)-Akt (Ser-473) (#9271), Akt1/2 (#9272), p-p44/42 MAPK (Erk1/2) (#9101) and Erk1/2 (#9102), cleaved-caspase-3 (#9664), cleaved-caspase-8 (#9496), cleaved-caspase-9 (#20750), cleaved-poly (ADP-ribose) polymerase (PARP) (#5625), β-tubulin (#15115) and β-actin (#3700) were purchased from Cell Signaling Tech (Beverly, MA).
Liu Z., Li P., Yang Y.Q., Cai S., Lin X., Chen M.B, & Guo H. (2020). I-BET726 suppresses human skin squamous cell carcinoma cell growth in vitro and in vivo. Cell Death & Disease, 11(5), 318.
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5

Western Blot Analysis of Protein Markers

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Cells were collected and total protein was extracted by RIPA lysis buffer (Beyotime, Shanghai, China, P0013 B), and BCA protein assay kit (Thermo Fisher Scientific, United States) was used to determine protein concentration. Equal amount of protein samples was resolved by SDS‐PAGE gel and the proteins were subsequently moved to PVDF membrane (Millipore, Bedford, MA), then the PVDF membrane were blocked (Beyotime, Shanghai, China, P0252) and immunoblotted with primary antibody of p44/42 MAPK (Erk1/2) (1:1,000; cat. no.4695T; Cell Signaling Technology), p-p44/42 MAPK (Erk1/2) (1:1,000; cat. no.4370T; Cell Signaling Technology), PD-L1 (1:1000, cat. no.66248-1-Ig, proteintech), GAPDH (1:1000, cat. no.60004-1-Ig, proteintech) at 4°C overnight. After washing by TBST (TBS containing 1% Tween), The membranes were exposed to corresponding secondary antibodies for 1 h. Antibody signal was detected using Clarity Western ECL substrate (Abbkine Scientific, China). The GAPDH served as an endogenous reference.
Zhang X., Wang K., Dai H., Cai J., Liu Y., Yin C., Wu J., Li X., Wu G., Lu A., Liu Q, & Guan D. (2022). Quantification of promoting efficiency and reducing toxicity of Traditional Chinese Medicine: A case study of the combination of Tripterygium wilfordii hook. f. and Lysimachia christinae hance in the treatment of lung cancer. Frontiers in Pharmacology, 13, 1018273.
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6

Potassium Oxoxazine Inhibits Urate-Induced Inflammation

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Potassium oxoxazine (Shanghai Macklin Biochemical Technology Co., Ltd C10097951, P831461), monosodium urate (Sigma U2875-5G), polyvinylidene fluoride (Millipore, USA.
， IPVH00005), α-tubulin (ABclonal, China, AC012), p44/42MAPK(ERK1/2) (Cell Signaling Technology, 4695 S), p-p44/42MAPK(ERK1/2) (Cell Signaling Technology, 4370 S), caspase-3(Cell Signaling Technology, ab184787）, Bcl-2 (Cell Signaling Technology, 3498 S), Bax (Abcam, UK, ab182733), U0126–EtOH (Med Chem Express, USA, HY-12031), HRP labeled goat anti-mouse IgG, HRP labeled goat anti-rabbit IgG, Alexa Fluor 488 labeled goat anti-rabbit IgG, Alexa Fluor 647 labeled goat anti-mouse IgG (Shanghai Biyuntian Biotechnology Co., Ltd, A0216, A0208, A0423, A0473)
Wu Z., Wang C., Yang F., Zhou J., Zhang X., Xin J, & Gao J. (2024). Network pharmacology, molecular docking, combined with experimental verification to explore the role and mechanism of shizhifang decoction in the treatment of hyperuricemia. Heliyon, 10(3), e24865.
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7

Western Blot Analysis of Protein Kinases

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Human FL-HCCs and normal liver samples were homogenized in ice-cold radioimmunoprecipitation buffer containing protease inhibitors, and protein concentrations were measured using the BCA Protein Assay (Pierce, Rockford, IL). Equal amounts of protein were separated by sodium dodecyl sulfate – poly-acrylamide gel electrophoresis (SDS-PAGE), transferred to Immobilin-P membranes (Millipore, Bedford, MA) and incubated at 4 °C overnight with the following primary antibodies: PKA C (SC-903, Santa Cruz Biotechnologies, Santa Cruz, CA), PKA RIIα (SC-909, Santa Cruz Biotechnologies), PKA RIIβ (610626, BD Biosciences, San Jose, CA), p-p44/42 MAPK (Erk1/2) (Thr202/Tyr204), total p44/42 MAPK (Erk1/2), and PKA RIα (9101, 9102, 5675, Cell Signaling, Danvers, MA), and β-Actin (A5441, Sigma, St. Louis, MO). Densitometry analysis was performed using NIH ImageJ software (National Institutes of Health, Bethesda, MD) by measuring the integrated density around each band and normalizing to the integrated density of the actin band for each sample.
Riggle K.M., Riehle K.J., Kenerson H.L., Turnham R., Homma M.K., Kazami M., Samelson B., Bauer R., McKnight G.S., Scott J.D, & Yeung R.S. (2016). Enhanced cAMP-stimulated protein kinase A activity in human fibrolamellar hepatocellular carcinoma. Pediatric research, 80(1), 110-118.
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8

Immunoblot Analysis of Signaling Pathways

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PTP1B activity was determined as previously described [35]. Cells or tissue were lysed in radio-immunoprecipitation assay (RIPA) buffer, centrifuged at 13,000 x g for 10 minutes and supernatants were collected. Immunoblots were conducted to determine levels of caspase-3, p-IκBα, IκBα (Ser32), p-p38 (Thr180/Tyr182), p38, TRIF, MyD88, IRAK1, p-p44/42 MAPK (ERK1/2) (Thr202/Tyr204), ERK1/2, β-actin (all antibodies from Cell Signaling Technologies, Beverly, MA) and S100A9 (Santa Cruz Biotechnologies).
Foronjy R.F., Ochieng P.O., Salathe M.A., Dabo A.J., Eden E., Baumlin N., Cummins N., Barik S., Campos M., Thorp E.B, & Geraghty P. (2016). Protein tyrosine phosphatase 1B negatively regulates S100A9-mediated lung damage during respiratory syncytial virus exacerbations. Mucosal immunology, 9(5), 1317-1329.
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9

Immunoblot Analysis of Signaling Pathways

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PTP1B activity was determined as previously described [35]. Cells or tissue were lysed in radio-immunoprecipitation assay (RIPA) buffer, centrifuged at 13,000 x g for 10 minutes and supernatants were collected. Immunoblots were conducted to determine levels of caspase-3, p-IκBα, IκBα (Ser32), p-p38 (Thr180/Tyr182), p38, TRIF, MyD88, IRAK1, p-p44/42 MAPK (ERK1/2) (Thr202/Tyr204), ERK1/2, β-actin (all antibodies from Cell Signaling Technologies, Beverly, MA) and S100A9 (Santa Cruz Biotechnologies).
Foronjy R.F., Ochieng P.O., Salathe M.A., Dabo A.J., Eden E., Baumlin N., Cummins N., Barik S., Campos M., Thorp E.B, & Geraghty P. (2016). Protein tyrosine phosphatase 1B negatively regulates S100A9-mediated lung damage during respiratory syncytial virus exacerbations. Mucosal immunology, 9(5), 1317-1329.
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10

Western Blot Protein Analysis Protocol

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Western blots were performed as previously described in Reference [26 (link)]. The primary antibodies used were: AR (1:1000; Santa Cruz Biotechnology), AR-V7 (1:400; Precision), GR (1:1000; BD transduction laboratories), PSA (1:1000; Santa Cruz Biotechnology), FKBP5 (1:1000; Santa Cruz Biotechnology), UBE2C (1:1000; Boston Biochem), NSE (1:1000; Merck), Mdr-1 (1:1000; Santa Cruz), Aurora A (1:1000), BRN-2 (1:1000), total-STAT3 (1:1000), p-STAT3Tyr705 (1:1000), total-AKT (1:1000), p-AktSer473 (1:1000), total-S6 (1:1000), p-S6 (1:2500), total-p44/42MAPKErk1/2 (1:1000), p-p44/42MAPKErk1/2 (1:1000), 110α (1:1000), 110β (1:1000), 110γ (1:1000), PI3KClass III; (1:1000), p85 (1:1000), 4EBP1 (1:1000), and p-4EBP1 (1:1000), from Cell Signaling Technology. Beta-actin (1:1000, Abcam and Cell Signaling Technology) was used as a loading control.
Shimizu Y., Tamada S., Kato M., Hirayama Y., Takeyama Y., Iguchi T., Sadar M.D, & Nakatani T. (2018). Androgen Receptor Splice Variant 7 Drives the Growth of Castration Resistant Prostate Cancer without Being Involved in the Efficacy of Taxane Chemotherapy. Journal of Clinical Medicine, 7(11), 444.
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