Erk1 2

Manufactured by Cell Signaling Technology

Sourced in United States, United Kingdom, China, Germany, Japan, Canada, France

ERK1/2 is a laboratory product that plays a core role in the extracellular signal-regulated kinase (ERK) pathway, a key signaling cascade involved in the regulation of various cellular processes. The product provides researchers with the necessary tools to study the activation and regulation of ERK1 and ERK2, two closely related kinases that are essential components of the ERK pathway.

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Lab products found in correlation

Close spelling variants of this product

Phospho-p44/42 MAPK (Erk1/2) (61 citations) Anti-phospho-ERK1/2 (Thr202/Tyr204) (57 citations) Phosphor-ERK1/2 (42 citations) Rabbit anti-p44/42 MAPK (Erk1/2) (23 citations) Rabbit anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (15 citations) Phospho-ERK1/2 (p-ERK1/2, (7 citations) Phos-ERK1/2 (7 citations) Antibodies against phospho-ERK1/2 (6 citations) PT202/Y204-ERK1/2 (6 citations) Anti-phospho-ERK1/2 #4370 (5 citations)

1 487 protocols using erk1 2

Immunoblotting and Immunofluorescence Antibody Protocol

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Immunoblot analyses were performed using the following antibodies: Akt, Aktp^S473, p70S6K, phospho-p70S6K, 4EBP1, ERK1/2, Atg5, ULK, ERK1/2, phopho-ERK1/2, p38, phospho-p38, phospho-NFκB and phospho-4EBP1 were all purchased from Cell Signaling. FAK (Santa Cruz Biotechnology, Inc.), actin (Cytoskeleton), LC3 (Novus) and FAKp^Y397 (BD Transduction Laboratories) were purchased from the suppliers indicated. Monoclonal anti-HA (16B12) and polyclonal anti-HA (HA.11) were both purchased from Covance. For immunofluorescence studies, LC3 (Millipore), mTOR (Cell Signaling), LAMP-1 and LAMP-2 (Developmental Studies Hybridoma Bank, University of Iowa), p62 (abcam) and ubiquitin (FK2, Enzo Life Sciences) were from the suppliers indicated. Secondary antibodies included goat anti-mouse-Cy3, donkey anti-rabbit Alexa Fluor 488 and goat anti-rat Alexa Fluor 555 and were purchased from Invitrogen. For flow cytometry, TLR4 (Sa15-21; Akashi et al., 2003) was conjugated to biotin and was a kind gift from Jonathan Kagan (Harvard Medical School, Boston, MA). Anti- strepavidin-APC was purchased from Biolegend.

Owen K.A., Meyer C.B., Bouton A.H, & Casanova J.E. (2014). Activation of Focal Adhesion Kinase by Salmonella Suppresses Autophagy via an Akt/mTOR Signaling Pathway and Promotes Bacterial Survival in Macrophages. PLoS Pathogens, 10(6), e1004159.

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Western Blot Analysis of Signaling Pathways

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Antibodies used were GSK3B (phospho S9) (9336 Cell Signalling), GSK3B (9315 Cell Signalling), ERK 1/2 (phospho T202, Y204) (9101 Cell Signalling), ERK 1/2 (9102 Cell Signalling), AKT (phospho S473) (9271 Cell Signalling), AKT (phospho T308) (9275 Cell Signalling), AKT (9272 Cell Signalling), JNK (phospho T183, Y195) (9251 Cell Signalling), JNK (9252 Cell Signalling), p38 (phospho T180, Y182) (9211 Cell Signalling), p38 (9212 Cell Signalling), B-ACTIN (A5441 Sigma), PEA15 (2780 Cell Signalling), PI3K (phospho Y458) (4228 Cell Signalling), PI3K (4229 Cell Signalling). PAK1/2 (phospho T423/T402) (2605 Cell Signalling), PAK1/2 (2604 Cell Signalling), Anti-rabbit IgG (A6154 Sigma and 7074 Cell Signalling) and anti-mouse IgG (7076 Cell Signalling) HRP-linked secondary antibodies were used for western blot detection.

Hunter J.E., Campbell A.E., Kerridge S., Fraser C., Hannaway N.L., Luli S., Ivanova I., Brownridge P.J., Coxhead J., Taylor L., Leary P., Hasoon M.S., Eyers C.E, & Perkins N.D. (2022). Up-regulation of the PI3K/AKT and RHO/RAC/PAK signalling pathways in CHK1 inhibitor resistant Eµ-Myc lymphoma cells. Biochemical Journal, 479(19), 2131-2151.

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Osthole Signaling Pathway Regulation

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Osthole (catalog number: O9265) was purchased from Sigma (St. Louis, MO, USA). Osthole was dissolved in DMSO to prepare a chemical stock for treatment. Antibodies against phosphorylated Akt (Ser⁴⁷³, catalog number: 4060), P70S6K (Thr⁴²¹/Ser⁴²⁴, catalog number: 9204), S6 (Ser²³⁵/Ser²³⁶, catalog number: 2211), ERK1/2 (Thr²⁰²/Tyr²⁰⁴, catalog number: 9101), p90RSK (Thr⁵⁷³, catalog number: 9346), JNK (Thr¹⁸³/Tyr¹⁸⁵, catalog number: 4668), total Akt (catalog number: 9272), P70S6K (catalog number: 9202), S6 (catalog number: 2217), ERK1/2 (catalog number: 4695), p90RSK (catalog number: 9335), JNK (catalog number: 9252), IRE1α (catalog number: 3294), eIF2α (catalog number: 5324), Bak (catalog number: 12105S), and Bax (catalog number: 2772) were purchased from Cell Signaling Technology (Beverly, MA, USA). Bcl-xL, p-Bcl-2, cleaved caspase 3 and cleaved caspase 9 were also purchased from cell Signaling Technology. Antibodies against GRP78 (catalog number: sc-13968), ATF6 α (catalog number: sc-166659), and α-tubulin (TUBA, catalog number: sc-32293) were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA). Inhibitors of ERK1/2 (U0126, catalog number: E1282) and JNK (SP600125, catalog number: E1305) were purchased from Enzo Life Sciences, Inc (Farmingdale, NY, USA), and a PI3K/Akt inhibitor (LY294002, catalog number: 9901) was purchased from Cell Signaling Technology, Inc.

Park W., Park S., Song G, & Lim W. (2019). Inhibitory Effects of Osthole on Human Breast Cancer Cell Progression via Induction of Cell Cycle Arrest, Mitochondrial Dysfunction, and ER Stress. Nutrients, 11(11), 2777.

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TGF-β1-Induced Fibroblast Signaling

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This analysis was performed with Pierce CoImmunoprecipitation (Co-IP) Kits (#26149, Pierce Biotechnology, Rockford, IL, USA) as previously described^{27 (link)}. Pulmonary fibroblasts from WT mice were treated with TGF-β1 and extracted for p16 (ab211542, Abcam), Bmi-1 (#6964, Cell Signaling Technology), ERK1/2 (#4695, Cell Signaling Technology) or pERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling Technology) coimmunoprecipitation analysis. Coprecipitates or total cytoplasmic lysates were detected with ERK1/2 (#4695, Cell Signaling Technology) and pERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling Technology) or p16 (ab211542, Abcam) antibodies for western blots analysis.

Chen H., Chen H., Liang J., Gu X., Zhou J., Xie C., Lv X., Wang R., Li Q., Mao Z., Sun H., Zuo G., Miao D, & Jin J. (2020). TGF-β1/IL-11/MEK/ERK signaling mediates senescence-associated pulmonary fibrosis in a stress-induced premature senescence model of Bmi-1 deficiency. Experimental & Molecular Medicine, 52(1), 130-151.

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Western Blot Analysis of Placental Proteins

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Western blot analysis was performed as described previously5 (link). Briefly, the placenta was homogenized to obtain protein samples. Then, the proteins (50 μg) were resolved by SDS-PAGE, transferred onto PVDF membranes, and probed with anti-phosphorylation antibodies to Akt (1:1000, Cell Signaling Technology, Inc., USA), mTOR (1:1000, Cell Signaling), p70S6K (1:1000, Bioworld Technology), ERK1/2 (1:1000, Cell Signaling) and antibody of TNF-α (1:500, Bioworld Technology) at 4 °C overnight. The membranes were then incubated with an HRP-labeled secondary antibody and developed using an ECL detection kit (Millipore, MA, USA). Following visualization, the blots were stripped by incubation in stripping buffer (Restore, Pierce) for 5 min, and then incubated with antibodies to Akt (1:1000, Cell Signaling), mTOR (1:1000, Cell Signaling), p70S6K (1:1000, Bioworld Technology), ERK1/2 (1:1000, Cell Signaling) and β-actin (1:1000, Cell Signaling). Western blot bands were scanned and analyzed with the National Institutes of Health Image image analysis software package.

Cao X., Hua X., Wang X, & Chen L. (2017). Exposure of pregnant mice to triclosan impairs placental development and nutrient transport. Scientific Reports, 7, 44803.

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Antibody-Based Investigation of Signaling Pathways

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Antibodies used in this study include rabbit antibodies against mGluR1a (Millipore; 1:2000), ERK1/2 (Cell Signaling; 1:2000), pERK1/2 (phosphorylation at T185/Y187 for pERK2; Cell Signaling; 1:1000), GST (Sigma; 1:2000), a phospho-serine/ threonine-proline motif (pS/TP, Abcam, Cambridge, MA; 1:1000), a phospho-MAPK motif PXpSP or pSPXR/K (Cell Signaling; 1:1000), phosphoserine (Invitrogen; 1:1000), phosphothreonine (Invitrogen; 1:1000), Src with phosphorylated tyrosine 416 (pY416, Cell Signaling; 1:1000), Src (Cell Signaling; 1:2000), or actin (Sigma; 1:4000) or mouse antibodies against mGluR1a (BD, Franklin Lakes, NJ; 1:2000), ERK1/2 (Cell Signaling; 1:2000), ERK2 (Abcam; 1:2000), or a phosphothreonine-proline motif (pTP, Cell Signaling; 1:1000). The anti-pY416 antibody reacts with the Src family members when phosphorylated at the conserved activation residue Y416. Pharmacological agents, including (S)-3,5-dihydroxyphenylglycine (DHPG), 3-methyl-aminothiophene dicarboxylic acid (3-MATIDA), and 1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio]butadiene (U0126), were purchased from Tocris Cookson Inc. (Ballwin, MO). All drugs were freshly prepared at the day of experiments.

Yang J.H., Mao L.M., Choe E.S, & Wang J.Q. (2016). Synaptic ERK2 Phosphorylates and Regulates Metabotropic Glutamate Receptor 1 In Vitro and in Neurons. Molecular neurobiology, 54(9), 7156-7170.

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Quantifying Kidney Fibrosis Markers

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The protein expression of CTGF, PAI‐1 (Santa Cruz Biotechnology, Heidelberg, Germany), Erk1/2, P‐Erk1/2 (Cell Signaling, Danvers, MA, USA) and TIMP‐1 (Sigma Aldrich, Taufkirchen, Germany) was assayed by western blotting 5. Representative immunoblots show the influence of UUO in comparison to the contralateral healthy kidney and the effects of BAY in healthy and fibrotic wt‐kidneys. The graphs statistically compare exclusive fibrotic kidneys. The values of all markers of the fibrotic wt‐ and cGKI‐KO‐kidneys were related to the mean values of fibrotic untreated wt‐kidneys. This ratio was set to one and normalized to the corresponding glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH; Cell Signaling) respectively to Erk1/2 for P‐Erk1/2 values. For quantification, ImageJ densitometry was used (BioRad, München, Germany).

Schinner E., Wetzl V., Schramm A., Kees F., Sandner P., Stasch J., Hofmann F, & Schlossmann J. (2017). Inhibition of the TGFβ signalling pathway by cGMP and cGMP‐dependent kinase I in renal fibrosis. FEBS Open Bio, 7(4), 550-561.

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Protein Isolation and Western Blot Analysis

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For the isolation of cellular proteins, cells were lysed in RIPA lysis buffer (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.5% NP-40, 15% Glycerol, supplemented with 10 µg/ml of each aprotinin, leupeptin and pepstatin as well as 0.8 µM Pefabloc (Roche, Mannheim, Germany), 1 mM NaF, and 1 mM Na₃VO₄). The protein concentration of the lysates was determined using Bradford Assay according to manufacturer´s instructions (Carl Roth, Karlsruhe, Germany). Proteins were separated by SDS-PAGE and transferred to a nitrocellulose membrane. Antigens were detected by incubation with specific primary antibodies (1:1,000) followed by incubation with DyLight-coupled secondary antibodies (1:10,000) (Licor, Lincoln, NE, USA). List of primary antibodies: (p)Y SHP2 (#3751), (p)Y STAT3 (#9145), STAT3 (#9139), (p)Y/T ERK1/2 (#4370), ERK1/2 (#4695) (Cell Signaling Technology); SOCS3 (#18391) (Immuno-Biological Laboratories, Fujioka, Japan); HSC70 (#7009) (Stress Marq, Victoria, Canada); SHP2 (#K0810) (Santa Cruz Biotechnology, Dallas, TEX, USA); Tubulin (#T5168) (Sigma-Aldrich Chemie, Munich, Germany). Detection was performed using an Odyssey gel documentation system (Licor). Analysis of Western blots was performed using Image Studio Lite (version 5.2, Licor).

Fiebelkow J., Guendel A., Guendel B., Mehwald N., Jetka T., Komorowski M., Waldherr S., Schaper F, & Dittrich A. (2021). The tyrosine phosphatase SHP2 increases robustness and information transfer within IL-6-induced JAK/STAT signalling. Cell Communication and Signaling : CCS, 19, 94.

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Hesperidin Modulation of Stress Signaling

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Hesperidin was purchased from Sigma-Aldrich and dissolved in dimethyl sulfoxide. Antibodies against phosphorylated AKT, P70S6K, ERK1/2, P90RSK, P38, eIF2α, and total AKT, P70S6K, ERK1/2, P90RSK, P38, IRE1α, and eIF2α were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibody against ATF6α was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). LY294002 was purchased from Cell Signaling Technology (Beverly, MA, USA). U0126 and SB203580 were purchased from Enzo Life Sciences (Farmingdale, NY, USA). Cisplatin and paclitaxel were purchased from Sigma-Aldrich (St.Louis, MO, USA).

Jeong S.A., Yang C., Song J., Song G., Jeong W, & Lim W. (2022). Hesperidin Suppresses the Proliferation of Prostate Cancer Cells by Inducing Oxidative Stress and Disrupting Ca2+ Homeostasis. Antioxidants, 11(9), 1633.

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Melatonin, Berberine, and Celecoxib Effects

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Melatonin, berberine and celecoxib were purchased from Sigma (St. Louis, MO) and dissolved in a small amount of DMSO before addition to the complete cell culture medium. Streptavidin-agarose was purchased from Sigma (St. Louis, MO). Antibodies to GAPDH, AP-2β, hTERT, COX-2, p50, cytochrome-c, PARP, caspase-9, BAX and Bcl-2 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies against Akt, ERK1/2, phosphrylated Akt or ERK1/2 were purchased from Cell Signaling (Beverly, MA).

Lu J.J., Fu L., Tang Z., Zhang C., Qin L., Wang J., Yu Z., Shi D., Xiao X., Xie F., Huang W, & Deng W. (2015). Melatonin inhibits AP-2β/hTERT, NF-κB/COX-2 and Akt/ERK and activates caspase/Cyto C signaling to enhance the antitumor activity of berberine in lung cancer cells. Oncotarget, 7(3), 2985-3001.

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