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Anti phospho mek1 2 ser217 221

Manufactured by Cell Signaling Technology
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Anti-phospho-MEK1/2 (Ser217/221) is a lab equipment product that detects the phosphorylation of MEK1 and MEK2 proteins at serine residues 217 and 221.

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Lab products found in correlation

Anti mek1 2, by Cell Signaling Technology (4 mentions) Anti erk1 2, by Cell Signaling Technology (3 mentions) Anti phospho erk1 2 thr202 tyr204, by Cell Signaling Technology (2 mentions) Anti stat1, by Cell Signaling Technology (1 mentions) Anti phospho stat1 tyr701, by Cell Signaling Technology (1 mentions) Anti β actin ab6276, by Abcam (1 mentions) Anti stat5 sc 835, by Santa Cruz Biotechnology (1 mentions) Newblot nitrocellulose stripping buffer, by LI COR (1 mentions) Anti phospho stat5 tyr694, by Cell Signaling Technology (1 mentions) Anti α tubulin, by Cell Signaling Technology (1 mentions) Mini protean precast gel, by Bio-Rad (1 mentions) Anti jak2, by Cell Signaling Technology (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Anti phospho raf, by Cell Signaling Technology (1 mentions) Anti mouse igg hrp, by Merck Group (1 mentions) Anti rabbit igg horseradish peroxidase hrp, by Cell Signaling Technology (1 mentions) Anti phospho pkcδ t505, by Cell Signaling Technology (1 mentions) Polyclonal rabbit anti active mapk, by Promega (1 mentions) Anti p70s6k, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Rabbit anti phospho-MEK1/2 (Ser217/221) Anti-phospho-MEK1/2(Ser217/221) antibody Phospho-MEK1/2 (Ser217/221) Anti-phospho-MEK1/2 Phospho-MEK1/2 Anti-MEK1/2 Phospho-MEK1 MEK1/2 Anti-MEK1

4 protocols using anti phospho mek1 2 ser217 221

1

Western Blot Analysis of JAK2 Signaling

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Leukemic cells were lysed in lysis buffer supplemented with freshly added protease and phosphatase inhibitors. 25μg (BCA method; Thermo Scientific) lysate was loaded on 10% mini protean precast gels (BioRad, Veenendaal, Netherlands), and transferred to a nitrocellulose membrane (Biorad). Primary antibody incubation was performed according to manufacturer's protocol. Anti-JAK2 (#3230), anti-phospho-JAK2-Tyr1007 (#4406), anti-phospho-STAT5-Tyr694 (#9351), anti-phospho-STAT1-Tyr701 (#9167), anti-Stat1 (#9175), anti-phospho-MEK1/2-Ser217/221 (#9154), anti-MEK1/2 (#4694), anti-phospho-Erk1/2-T202/204 (#4370), anti-Erk1/2 (#91078), and anti-αTubulin (#2144) were obtained from Cell Signaling Technology (Danvers, Massachusetts, USA). Anti-β-actin (ab6276) was obtained from Abcam (Cambridge, UK), and anti-STAT5 (sc-835) from Santa Cruz (Heidelberg, Germany). Blots were stained with secondary antibodies (IRDye 680RD- or 800CW-labelled anti-rabbit and IRDye 680RD- or 800CW-labelled anti-mouse; Li-Cor Biosciences, Leusden, Netherlands) and scanned using the Odyssey infrared imaging system (Li-Cor Biosciences). To reprobe membranes, they were stripped in NewBlot Nitrocellulose stripping buffer (Li-Cor Biosciences) according to manufacturer's protocol. BCR-JAK2, PAX5-JAK2 and TERF2-JAK2 proteins were separated from wildtype JAK2 based on size (~94, 57, 95 and 125 kDa, respectively).
Steeghs E.M., Jerchel I.S., de Goffau-Nobel W., Hoogkamer A.Q., Boer J.M., Boeree A., van de Ven C., Koudijs M.J., Besselink N.J., de Groot-Kruseman H.A., Zwaan C.M., Horstmann M.A., Pieters R, & den Boer M.L. (2017). JAK2 aberrations in childhood B-cell precursor acute lymphoblastic leukemia. Oncotarget, 8(52), 89923-89938.
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2

Immunoblotting of Raf-MEK-ERK Pathway

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Purified CD4+ T cells were suspended in RPMI 1640 supplemented with 10% FCS, 2 mM glutamine and penicillin/streptomycin then left unstimulated or stimulated with 50 ng/ml PMA for 15 min at 37°C. Where indicated, cells were incubated in the presence or absence of Hyd (10 μM), H2O2 (50 μM), or ONOO− (50 μM) for 60 min before PMA stimulation. Following stimulation the cells were lysed, then total and phosphorylated ERK, MEK, RAF and PKCδ measured by immunoblotting as described (16 (link)). The following primary Abs were used at a 1/1000 dilution: anti-phospho-raf (Ser338), anti-phospho-MEK1/2 (Ser217/221), anti-MEK1/2 and anti-phospho-PKCδ (T505) (Cell Signaling Technology, Beverly MA). Polyclonal rabbit anti-active MAPK (1/5000) was purchased from Promega. Secondary Abs included anti-rabbit IgG horse radish peroxidase (HRP) (1/2000; Cell Signaling Technology) and anti-mouse IgG HRP (1/4000; Sigma-Aldrich).
Li Y., Gorelik G., Strickland F.M, & Richardson B.C. (2014). Oxidative Stress, T Cell DNA Methylation, and Lupus. Arthritis & Rheumatology (Hoboken, N.j.), 66(6), 1574-1582.
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3

Bilirubin and MAPK Pathway Inhibitors

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Bilirubin (B4126-5G) was purchased from Sigma-Aldrich (USA). Vemurafenib (S1267), KO-947 (S8569), and LY3214996 (S8534) were obtained from Selleckchem (Houston, TX). Antibodies, including anti-PARP (#9532), Caspase 3 (#9662), anti-cleaved Caspase-3 (Asp175) (#9661), anti-Caspase 9 (#9502), anti-cleaved Caspase-9 (Asp315) (#9505), anti-phospho-B-Raf (Ser445) (#2696), anti-B-Raf (#14814), anti-phospho-MEK1/2 (Ser217/221) (#3958), anti-MEK1/2 (#4694), anti-phospho-ERK1/2 (Thr202/Tyr204) (#4370), anti-ERK1/2 (#4695), anti-phospho-MNK1 (Thr197/202) (#2111), anti-MNK1 (#2195), anti-phospho-eIF4E (Ser209) (#9741), anti-eIF4E (#2067), anti-phospho-p70S6K (Thr421/Ser424) (#9204), anti-p70S6K (#2708), anti-phospho-4eBP1(Thr37/46) (#2855), anti-4eBP1 (#9644), and anti-GAPDH (#5174) were purchased from Cell Signaling Technology (Beverly, MA).
Tan Y., Zhong X., Wen X., Yao L., Shao Z., Sun W., Wu J., Wen G., Tang D., Zhang X., Liao Y, & Liu J. (2021). Bilirubin Restrains the Anticancer Effect of Vemurafenib on BRAF-Mutant Melanoma Cells Through ERK-MNK1 Signaling. Frontiers in Oncology, 11, 698888.
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4

Western Blot Analysis of Signaling Proteins

Cited 1 time
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Western blot analysis was carried out as previously described [30 (link)]. The following primary antibodies (Abs) were used: anti-phospho-H2A.X (S139) (1/2000), anti-H2A.X (1/1000), anti-phospho-ERK1/2 (Thr202/Tyr204) (1/2000), anti-ERK1/2 (1/2000), anti-phospho-MEK1/2 (Ser217/221) (1/2000), and anti-MEK1/2 (1/2000) from Cell Signaling Technology (Danvers, MA, USA), and anti-PARP-1 (1/200; Santa Cruz Biotechnology, (Dallas, TX, USA)) and anti-Actin (1/10,000; Sigma-Aldrich). The following secondary Abs were used: anti-rabbit-HRP (1/5000) and anti-mouse-HRP (1/5000 and 1/10,000 for actin) from SouthernBiotech (Nanterre, France). The Western blots were scanned using ChemiDoc imaging system (Bio-Rad, Marnes-la-Coquette, France). The densitometric analyses were performed using an ImageJ software (National Institutes of Health, Bethesda, MD, USA) (accessed on 11 November 2020).
Akil H., Quintana M., Raymond J.H., Billoux T., Benboubker V., Besse S., Auzeloux P., Delmas V., Petit V., Larue L., D’Incan M., Degoul F, & Rouanet J. (2021). Efficacy of Targeted Radionuclide Therapy Using [131I]ICF01012 in 3D Pigmented BRAF- and NRAS-Mutant Melanoma Models and In Vivo NRAS-Mutant Melanoma. Cancers, 13(6), 1421.
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