- PA5-29636 - Invitrogen Antibodies MAPK1 antibody

Hao Y, Wang Z, Wang X, Zhan W, Wu D
Journal of thoracic disease 2021 Aug;13(8):5042-5054

Zhu D, Shao M, Yang J, Fang M, Liu S, Lou D, Gao R, Liu Y, Li A, Lv Y, Mo Z, Fan Q
Journal of Cancer 2020;11(8):2360-2370

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (1), H1975 (2), HCT 116 (3), HT-29 (4), A-431 (5), A549 (6), MDA-MB-231 (7) and MCF7 (8). The blot was probed with Rabbit Anti-ERK2 Polyclonal Antibody (Product # PA5-29636, 1:5000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 42 kDa band corresponding to ERK2 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of ERK2 was achieved by transfecting MCF7 cells with ERK2 specific validated siRNAs (Silencer® select Product # s11137 and s11138). Western Blot analysis (Fig a) was performed using whole cell lysates from the ERK2 knockdown cells (3), ERK1 siRNA transfected cells (4), ERK1/2 siRNA transfected cells (5), non-specific scrambled siRNA transfected cells (2) and untransfected cells (1). The blots were probed with Anti-ERK2 Rabbit Polyclonal Antibody (Product # PA5-29636, 1 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Decrease in signal upon siRNA mediated knockdown confirms that the antibody is specific to ERK2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using ERK2 Polyclonal Antibody (Product # PA5-29636). Sample (50 µg of whole cell lysate). Lane A: mouse brain. 10% SDS PAGE. ERK2 Polyclonal Antibody (Product # PA5-29636) diluted at 1:5,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using ERK2 Polyclonal Antibody (Product # PA5-29636). Sample (30 µg of whole cell lysate). Lane A: H1299. Lane B: Hela. 10% SDS PAGE. ERK2 Polyclonal Antibody (Product # PA5-29636) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: ERK2 Polyclonal Antibody detects ERK2 protein at cytosol and nucleus on human breast carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. ERK2 Polyclonal Antibody (Product # PA5-29636) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: ERK2 Polyclonal Antibody detects ERK2 protein at cytosol and nucleus on human colon carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human colon carcinoma. ERK2 Polyclonal Antibody (Product # PA5-29636) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: ERK2 Polyclonal Antibody immunoprecipitates MAPK1 protein in IP experiments. IP samples: HeLa whole cell extract. A. 30 µg HeLa whole cell extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of MAPK1 protein by 4 µg ERK2 Polyclonal Antibody (Product # PA5-29636). 10 % SDS-PAGE. The immunoprecipitated MAPK1 protein was detected by ERK2 Polyclonal Antibody (Product # PA5-29636) diluted at 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Verification of the circRNA-miRNA-mRNA interaction network. (A1) qRT-PCR detection of circRNA expression in the three groups. (A2) Levels of target miRNAs for circRNA in the three groups. (A3) Levels of target mRNAs for miRNAs in the three groups. The expression of each RNA was calculated using 2-^^Ct equation. Compared to NC group: *p < 0.05, and compared to IM group: #p < 0.05. (B) WB analysis of selected protein expression. Compared to NC group: *p < 0.05, and compared to IM group: #p < 0.05.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 MAPK is the downstream pathway of OGDH. (A) According to the expression data of GSE16650, the KEGG analysis was used to generate the bubble plot; (B) GSEA analysis showed that the gene set 'KEGG_MAPK_SIGNALING_pathway' was highly expressed in the high expression group of OGDH; (C) in the CLP animal model, down-regulation of OGDH reversed the increase of p-MAPK1 expression; (D) in an LPS-induced lung injury cell model, down-regulation of OGDH can down-regulate p-MAPK1 expression; (E) immunofluorescence showed down-regulation of OGDH and down-regulation of p-MAPK1 expression. OGDH, oxoglutarate dehydrogenase.

PA5-29636