Myc clone 9e10

Manufactured by Merck Group

Sourced in United States

The MYC (clone 9E10) is a laboratory reagent produced by Merck Group. It is a monoclonal antibody that recognizes the MYC protein, a transcription factor involved in cellular processes. The antibody can be used for the detection and study of the MYC protein in various applications, such as Western blotting, immunoprecipitation, and immunohistochemistry. The core function of this product is to serve as a specific tool for researchers investigating the MYC protein and its role in biological systems.

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Lab products found in correlation

Flag clone m2, by Merck Group (2 mentions) Goat anti mouse igg hrp, by Santa Cruz Biotechnology (2 mentions) Ha clone 12ca5, by Roche (1 mentions) Gapdh clone 6c5, by Abcam (1 mentions) Flotillin 2, by BD (1 mentions) Rabbit anti ubc9, by Santa Cruz Biotechnology (1 mentions) α synuclein, by Thermo Fisher Scientific (1 mentions) Rat anti lamp1 1d4b, by Santa Cruz Biotechnology (1 mentions) Rabbit anti gfp, by Thermo Fisher Scientific (1 mentions) Tsg101, by GeneTex (1 mentions) R960 25, by Thermo Fisher Scientific (1 mentions) Sc9071, by Santa Cruz Biotechnology (1 mentions) Cab1001, by Thermo Fisher Scientific (1 mentions) Image lab software, by Bio-Rad (1 mentions) Chemidoc imaging system, by Bio-Rad (1 mentions) Gardiquimod, by InvivoGen (1 mentions) Mouse anti rabbit igg hrp, by Santa Cruz Biotechnology (1 mentions) Ifnα2, by Merck Group (1 mentions) Tautomycin, by Merck Group (1 mentions)

4 protocols using myc clone 9e10

Immunoprecipitation and Western Blot Protocol

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Transfected cells were lysed in Nonidet P-40-desoxycholate buffer (50 mM Tris pH 7.4, 150 mM NaCl, 1% Nonidet P-40, 0.5% Na desoxycholate, 0.5 mM Tris(2-carboxyethyl)phosphine, and 0.5 mM Pefabloc). Immunoprecipitation were carried out by addition of antibodies diluted 1:250 and incubation at 4 °C for 1h30. Protein A sepharose beads were added and incubated with the mix for 1 h. Beads were collected by centrifugation and washed three times with Nonidet P-40-desoxycholate buffer. For experience in Supplementary Fig. 1 protein G magnetic beads were used. Proteins were eluted in 2× SDS sample buffer at 80 °C for 10 min. After protein gel electrophoresis and transfer to PVDF membrane, detection was carried out by incubation with primary antibodies diluted 1:1000 or as indicated by the manufacturer and revelation was performed by chemiluminescence using ECL, ECL + or ECL Prime kits. Following antibodies were purchased: FLAG (clone M2, Sigma), MYC (clone 9E10, Sigma), HA (clone 12CA5, Roche), SMAD3 (NB600-1258), SMAD2/3 (566412, Calbiochem) and TAL1 (BTL73, Millipore).

Terme J.M., Lemaire S., Auboeuf D., Mocquet V, & Jalinot P. (2016). The proto-oncogenic protein TAL1 controls TGF-β1 signaling through interaction with SMAD3. Biochimie Open, 2, 69-78.

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Antibody Profiling of Cellular Organelles

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Primary antibodies were: mouse monoclonal antibodies against Myc clone 9E10 (Sigma), Flotillin-2 (BD Biosciences), α-Synuclein (Invitrogen), Alix (BD Biosciences), TSG101 (GeneTex Inc., Irvine, CA, USA), CD63 (BD Biosciences), 6E10 anti-APP (Signet), GAPDH clone 6C5 (Abcam), rabbit anti-Glutamate Receptor GluR2/3 (Chemicon), rabbit anti-Glutamate Receptor GluR1 (Chemicon), rabbit anti-Calnexin (StressGene), rabbit anti-GFP (Invitrogen), rabbit anti-Integrin β5 (Millipore), rabbit anti-UBC9 (Santa Cruz), and rat anti-LAMP1 (1D4B) (Santa Cruz). SUMO-2 antibody was kindly provided by F. Melchior (Heidelberg, ZMBH, Germany) [5 (link)]. Secondary antibodies were obtained from Dianova and Invitrogen.

Kunadt M., Eckermann K., Stuendl A., Gong J., Russo B., Strauss K., Rai S., Kügler S., Falomir Lockhart L., Schwalbe M., Krumova P., Oliveira L.M., Bähr M., Möbius W., Levin J., Giese A., Kruse N., Mollenhauer B., Geiss-Friedlander R., Ludolph A.C., Freischmidt A., Feiler M.S., Danzer K.M., Zweckstetter M., Jovin T.M., Simons M., Weishaupt J.H, & Schneider A. (2015). Extracellular vesicle sorting of α-Synuclein is regulated by sumoylation. Acta Neuropathologica, 129(5), 695-713.

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Western Blot Analysis of Cell Lysates

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A total of 1 OD cell pellets were processed using TCA lysis and Western blots performed as previously described (Leech et al. 2020 (link)). For non-reducing lysis, lysates were prepared the same way except that 4X SDS-PAGE sample buffer without β-mercaptoethanol (40% glycerol, 0.25 M Tris pH 6.8, 8% SDS, bromophenol blue) was used. Western blotting was performed with antibodies against Clb2 (sc-9071, Santa Cruz), FLAG (clone M2, F1804, Sigma), G6PDH (A9521, Sigma), MYC (clone 9E10, M5546, Sigma), PSTAIRE (P7962, Sigma), TAP (CAB1001, ThermoFisher), and V5 (R960-25, ThermoFisher). The PSTAIRE antibody recognizes a conserved sequence in CDK kinases and is used as a loading control.
To quantify Western blot data, images were collected on a ChemiDoc Imaging System (BioRad) and quantified using Image Lab software (Biorad). For all experiments, a minimum of three biological replicates was performed, with the exact number noted in the figure legends. Data were analyzed using GraphPad Prism software.

Arsenault H.E., Ghizzoni J.M., Leech C.M., Diers A.R., Gesta S., Vishnudas V.K., Narain N.R., Sarangarajan R, & Benanti J.A. (2021). Ubc1 turnover contributes to the spindle assembly checkpoint in Saccharomyces cerevisiae. G3: Genes|Genomes|Genetics, 11(12), jkab346.

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Plasmid Constructs for PP1, IRF7, and TLR7 Studies

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Flag-PP1 (α, β and γ) expression plasmids cloned in Flag-CMV4 were provided by Dr. Sherry Winter [53 (link)]. Flag-IRF7 expression plasmids and its mutants were described previously [54 , 55 (link)]. 2XMyc-IRF7 plasmids were generated by subcloning of 2XMyc-IRF7 fragment (by PCR amplification from pcDNA3-IRF7) into pCMV-Tag3B (Stratagene). TLR7 cDNA was cloned from human genome (the human B cell line BJAB) by PCR amplification. Deletion and point mutants were generated by site-directed mutation (Stratagene), and verified by sequencing. PE-conjugated IRF7(p477/479) antibody (clone K47–671) for flow cytometry was purchased from BD Biosciences. The same clone with higher concentration but without conjugation was customized for immunoblotting. Mouse anti-PP1 (clone E-9), mouse anti-PP1α (clone G-4), goat anti-PP1α (clone C19), mouse anti-IRF7 (clone G-8), rabbit anti-IRF7 (clone H-246), goat anti-mouse IgG-HRP, mouse anti-rabbit IgG-HRP, and mouse anti-goat IgG-HRP, were purchased from Santa Cruz. Flag (clone M2) and Myc (clone 9E10) antibodies were from Sigma and Roche, respectively. FITC-, PE-, APC-, and PerCP-eFluor710-conjugated antibodies for flow cytometry were purchased from eBioscience. Gardiquimod (designated as “Gardi” in figure legend) and HIV-1 ssRNA40 and controls were purchased from Invivogen. IFNα2 was purchased from Sigma. Tautomycin was purchased from EMD Millipore.

Wang L., Zhao J., Ren J., Hall K.H., Moorman J.P., Yao Z.Q, & Ning S. (2016). Protein phosphatase 1 abrogates IRF7-mediated type I IFN response in antiviral immunity. European journal of immunology, 46(10), 2409-2419.

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