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Anti phospho erk1 2 d13.14.4e

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Anti-phospho ERK1/2 (D13.14.4E) is a rabbit monoclonal antibody that specifically recognizes the phosphorylated forms of extracellular signal-regulated kinase 1 and 2 (ERK1/2).

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Lab products found in correlation

Goat anti rabbit igg, by LI COR (2 mentions) Anti erk1 2 137f5, by Cell Signaling Technology (2 mentions) Goat anti mouse igg conjugated to irdye800cw, by LI COR (2 mentions) Anti actin antibody, by Merck Group (1 mentions) Anti actin antibody, by Thermo Fisher Scientific (1 mentions) Anti phospho p38 d3f9, by Cell Signaling Technology (1 mentions) Anti α tubulin dm1a, by Merck Group (1 mentions) Anti phospho cofilin ser3, by Cell Signaling Technology (1 mentions) Anti yap sc 101199, by Santa Cruz Biotechnology (1 mentions) Anti erk1 2, by Cell Signaling Technology (1 mentions) Phytohemagglutinin (pha), by Merck Group (1 mentions) Annexin v 7 aad apoptosis detection kit, by BD (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Anti cd4 gk1, by Thermo Fisher Scientific (1 mentions) Indo 1 am, by Thermo Fisher Scientific (1 mentions) Anti human cd28 cd28.2, by Thermo Fisher Scientific (1 mentions) Anti β actin ac 74, by Merck Group (1 mentions) Fura 2 am, by Thermo Fisher Scientific (1 mentions) Thapsigargin, by Merck Group (1 mentions)

Spelling variants of this product

ERK1/2 (1487 citations) Anti-ERK1/2 (692 citations) Phospho-ERK1/2 (582 citations) Anti-phospho-ERK1/2 (429 citations) Anti-phospho-p44/42 MAPK (Erk1/2)(Thr202/Tyr204) (D13.14.4E) (4 citations) Phospho-ERK1/2 (D13.14.4E), (3 citations)

4 protocols using anti phospho erk1 2 d13.14.4e

1

Western Blot Analysis of Signaling Proteins

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After the indicated treatments, the cells were washed three times with PBS. The cell lysates were prepared via the addition of SDS-PAGE sample reducing buffer, followed by heating at 95°C for 10 min. Immunoblotting was performed using anti-phospho ERK1/2 (D13.14.4E) (1:1,000 dilution), anti-ERK1/2 (137F5) (1:1,000 dilution) and anti-IκBα alpha (L35A5) (1:2,000 dilution) antibodies from Cell Signaling Technology. An anti-actin antibody (Millipore) (1:5,000 dilution) was used to confirm equal loading. The secondary antibodies included goat anti-rabbit IgG and goat anti-mouse IgG conjugated to IRdye800CW (Li-COR) (1:10,000 dilution). ImageJ software was used to quantify band intensities, and actin was used to normalize the total amount of protein loaded in each well.
Tavares R, & Pathak S.K. (2015). Helicobacter pylori Protein JHP0290 Exhibits Proliferative and Anti-Apoptotic Effects in Gastric Epithelial Cells. PLoS ONE, 10(4), e0124407.
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2

Cell Lysate Preparation and Immunoblotting

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The method used for cell lysate preparation and immunoblotting were described previously (Pathak et al., 2013 (link)). Briefly, cell lysates were prepared by the addition of SDS-PAGE sample reducing buffer to cells after washing twice with PBS, followed by heating at 95°C for 10 min. Immunoblotting was performed using anti-phospho ERK1/2 (D13.14.4E) (1:1,000 dilution), anti-phospho-p38 (D3F9), anti-phospho-JNK (81E11), and anti-ERK1/2 (137F5) (1:1,000 dilution) antibodies from Cell Signaling Technology. Anti-phospho-c-Fos (Thr325) antibody was from Bioss antibodies. An anti-actin antibody (Thermo Scientific) (1:5,000 dilution) was used to confirm equal loading. The secondary antibodies included goat anti-rabbit IgG and goat anti-mouse IgG conjugated to IRdye800CW (Li-COR) (1:10,000 dilution). Band intensities were quantified using the ImageJ software, and actin was used to normalize the total amount of protein loaded in each well.
Tavares R, & Pathak S.K. (2017). Helicobacter pylori Secreted Protein HP1286 Triggers Apoptosis in Macrophages via TNF-Independent and ERK MAPK-Dependent Pathways. Frontiers in Cellular and Infection Microbiology, 7, 58.
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3

Antibody Immunoblot Analysis

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The antibodies in this study included anti-α-tubulin (DM1A; Sigma), anti-Erk1/2 (137F5; Cell Signaling Technology), anti-Phospho-Erk1/2 (D13.14.4E; Cell Signaling Technology), anti-Cofilin (ACFL02; Cytoskeleton), anti-Phospho-Cofilin (Ser3) (#3311; Cell Signaling Technology), and anti-YAP (sc101199; Santa Cruz) (detects both YAP and TAZ).
Ito T., Taniguchi H., Fukagai K., Okamuro S, & Kobayashi A. (2015). Inhibitory Mechanism of FAT4 Gene Expression in Response to Actin Dynamics during Src-Induced Carcinogenesis. PLoS ONE, 10(2), e0118336.
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4

Immunoblotting and Immunofluorescence Analysis

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For immunoblotting, we used: anti-phospho Erk1/2 (D13.14.4E), p38 (D3F9), and Jnk (81E11), and anti-total Erk1/2 (137F5), p38 (#9212), Jnk (#9252) Abs; anti-phospho Zap70 (Tyr319) and PLC-γ1 (Tyr783) Abs (Cell Signaling Technology), anti-phospho Lat (Tyr191) and anti-p-Tyr (4G-10) Abs (Millipore); Anti-CD3ε (M-20; sc-1127), anti-Lck (2102; sc-13) and anti-TRPV1 (P-19; sc-12498) Abs (Santa Cruz Biotechnology); anti-TRPV1 (#ACC-030, Alomone), anti-NFAT1 mAb (ab2722, Abcam), and anti-β-actin (AC-74, Sigma). For immunofluorescence studies, anti-TRPV1 (sc-12498) and anti-Lck (sc-13) (Santa Cruz Biotechnology), anti-CD4 (GK1.5, eBioscience), anti-CD45 and anti-Claudin-3 Abs (BD Biosciences). For T cell stimulation, anti-mouse CD3ε (145-2c11) and anti-mouse CD28 (PV-1) (BioXcell), and anti-human CD3ε (UCHT1) and anti-human CD28 (CD28.2) (eBioscience) mAbs were used; PMA (Phorbol 12-myristate 13-acetate), ionomycin, thapsigargin and phytohemagglutinin were from Sigma; The Src-family kinase inhibitor PP2 was from Cayman Chemical, the TRPV1 antagonists were from Tocris (BCTC, I-RTX) and Enzo Life Sciences (SB366791), the Annexin-V/7-AAD apoptosis detection kit was from BD Biosciences and CFSE, Indo-1 AM, and Fura-2 AM were from Invitrogen/Molecular Probes.
Bertin S., Aoki-Nonaka Y., de Jong P.R., Nohara L.L., Xu H., Stanwood S.R., Srikanth S., Lee J., To K., Abramson L., Yu T., Han T., Touma R., Li X., González-Navajas J.M., Herdman S., Corr M., Fu G., Dong H., Gwack Y., Franco A., Jefferies W.A, & Raz E. (2014). The ion channel TRPV1 regulates the activation and proinflammatory properties of CD4+ T cells. Nature immunology, 15(11), 1055-1063.
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