Myc tag 9b11

Manufactured by Cell Signaling Technology

Sourced in United States

Myc-Tag (9B11) is a monoclonal antibody that recognizes the Myc epitope tag. The Myc tag is a short peptide sequence (EQKLISEEDL) derived from the c-Myc protein, commonly used for the detection and purification of recombinant proteins in various applications.

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Lab products found in correlation

Hoechst 33342, by Thermo Fisher Scientific (3 mentions) Anti lamin b1, by Abcam (2 mentions) Anti vdac1, by Abcam (2 mentions) Mouse trueblot ultra anti mouse ig hrp, by Rockland Immunochemicals (2 mentions) Caspase 3, by Cell Signaling Technology (2 mentions) Anti phospho histone h2a x ser139, by Merck Group (2 mentions) α tubulin 4g1, by Santa Cruz Biotechnology (2 mentions) Bap1 c 4, by Santa Cruz Biotechnology (2 mentions) Bap1 d7w7o, by Cell Signaling Technology (2 mentions) Bap1 h 300, by Santa Cruz Biotechnology (2 mentions) Anti human cd90, by BD (2 mentions) Histone h2a x d17a3, by Cell Signaling Technology (2 mentions) Ip3r1, by Novus Biologicals (2 mentions) C myc, by Fortis Life Sciences (2 mentions) Goat anti mouse igg h, by Thermo Fisher Scientific (2 mentions) Goat anti rabbit igg h, by Thermo Fisher Scientific (2 mentions) Flag tag (flag), by Merck Group (2 mentions) Ip3r3, by BD (2 mentions) Anti flag m2, by Merck Group (2 mentions) Lsm 510, by Zeiss (2 mentions)

15 protocols using myc tag 9b11

Comprehensive Antibody Panel for Western Blotting

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Antibodies used for Western blotting: YAP [D8H1X (#14074) or 1A12 (#12395)], TAZ [D3I6D (#70148) or E5P2N (#71192)], Myc-tag [9B11 (#2276) or 71D10 (#2278)], panTEAD [D3F7L (#13295)], Rap1-interacting factor 1 (RIF1) [D2F2M (#95558)], GAPDH [D16H11 (#5174)], α-Tubulin [DM1A (#3873)], pS139 H2A.X [20E3 (#9718)], Ku70 [D10A7 (#4588)], and Ku80 (#2753) from Cell Signaling Technology. MAX [(H2) sc-8011] from Santa Cruz Biotechnology. Secondary antibodies: Goat anti-mouse (926–3220, Li-cor) and Goat-anti-rabbit (926–68071, Li-Cor).
Antibody used for immunoprecipitation: Myc-tag [9B11 (#2276) or 71D10 (#2278), Cell Signaling Technology], Mouse (G3A1) mAb IgG1 Isotype Control (#5415, Cell Signaling Technology).
Antibodies used for immunofluorescence: panTEAD [D3F7L (#13295), Cell Signaling Technologies], RIF1 (NBP2-26219, Novus biologicals), pS139 H2A.X (05–636, Millipore-Sigma).

Calses P.C., Pham V.C., Guarnaccia A.D., Choi M., Verschueren E., Bakker S.T., Pham T.H., Hinkle T., Liu C., Chang M.T., Kljavin N., Bakalarski C., Haley B., Zou J., Yan C., Song X., Lin X., Rowntree R., Ashworth A., Dey A, & Lill J.R. (2023). TEAD Proteins Associate With DNA Repair Proteins to Facilitate Cellular Recovery From DNA Damage. Molecular & Cellular Proteomics : MCP, 22(2), 100496.

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Immunostaining and Stress Granule Analysis

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Cells grown on glass coverslips were fixed and immunostained according to the protocol in [6] (link) using mouse monoclonal antibodies to IAV nucleoprotein (AAH5; Abcam), G3BP (clone 23, BD Transduction Labs), and PABP1 (sc-32318, Santa Cruz Biotechnology); goat polyclonal antibody to TIA-1 (sc-1751, Santa Cruz Biotechnology), influenza virus (ab20841, Abcam), rabbit antibody to TIAR (clone D32D3, Cell Signaling), YB-1 (ab12148, Abcam) and myc-tag (9B11; Cell Signaling) at manufacturer-recommended dilutions. AlexaFluor-conjugated secondary antibodies (Molecular Probes) were used at 1∶1000 dilution. Images were captured using Zeiss Axioplan II microscope or Zeiss LSM 510 laser scanning microscope. Quantification of stress granules was done in at least 3 random fields of view with greater than 200 cells analysed on each slide. Cells were considered stress granule-positive if two or more stress granule marker foci were present in the cytoplasm. For western blot analysis, whole cell lysates were resolved on denaturing 10% polyacrylamide gels and analyzed using primary antibodies described above and the antibodies to phospho-Ser-51- eIF2α (rabbit, D9G8, Cell Signaling), eIF4A (goat, sc-14211, Santa Cruz Biotechnology), and actin (rabbit, 4968; Cell Signaling).

Khaperskyy D.A., Emara M.M., Johnston B.P., Anderson P., Hatchette T.F, & McCormick C. (2014). Influenza A Virus Host Shutoff Disables Antiviral Stress-Induced Translation Arrest. PLoS Pathogens, 10(7), e1004217.

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Comprehensive Antibody Characterization Protocol

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Primary antibodies used were: α-Tubulin (4G1) (Santa Cruz Biotechnology, cat. no. sc-58666), BAP1 (C-4) (Santa Cruz Biotechnology, cat. no. sc-28383), BAP1 (D7W7O) (Cell Signaling, cat. no. 13271), BAP1 (H-300) (Santa Cruz Biotechnology, cat. no. sc-28236), anti-human CD90 (BD Biosciences, cat. no. 561558), Caspase-3 (Cell Signaling, cat. no. 9662), FLAG (Sigma-Aldrich, cat. no. F7425), Histone H2A.X (D17A3) (Cell Signaling, cat. no. 7631), Anti-phospho-Histone H2A.X (Ser139) (EMD Millipore, cat. no. 05-636), H3 (Cell Signaling, cat. no. 4499), IP3R1 (Novus Biologicals, cat. no. NB120-5908), IP3R3 (BD Biosciences, cat. no. 610312), Anti-Lamin B1 (abcam, cat. no. ab16048), c-myc (Bethyl Laboratories, cat. no A190-105A), Myc-Tag (9B11) (Cell Signaling, cat. no. 2276), PDI (abcam, cat. no. ab31811) Anti-VDAC1 (abcam, cat. no. ab15895). Secondary antibodies used were: Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP (Thermo Scientific, cat. no. 32430); Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP (Thermo Scientific, cat. no. 32460); Mouse TrueBlot® ULTRA: Anti-Mouse Ig HRP (Rockland, cat. no. 18-8817-31).

Bononi A., Giorgi C., Patergnani S., Larson D., Verbruggen K., Tanji M., Pellegrini L., Signorato V., Olivetto F., Pastorino S., Nasu M., Napolitano A., Gaudino G., Morris P., Sakamoto G., Ferris L.K., Danese A., Raimondi A., Tacchetti C., Kuchay S., Pass H.I., Affar E.B., Yang H., Pinton P, & Carbone M. (2017). BAP1 regulates IP3R3-mediated Ca2+ flux to mitochondria suppressing cell transformation. Nature, 546(7659), 549-553.

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Immunoblotting Antibody Validation Protocol

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Caveolin-1 (pAB #610060, 2234 #610494 and 2297 #610406), caveolin-2 (#610684) and GM130 (#610822) antibodies were purchased from BD Biosciences (Table 2). Caveolin-1 C-term [E249] (ab32577), PTRF/Cavin1 (ab48824) and EHD2 (ab23935) antibodies were purchased from Abcam. The PACSIN2 (AP088b) antibody used was purchased from Abgent. Alexa-fluor dye-conjugated secondary antibodies were purchased from Life Technologies. A c-Myc (A-14) antibody was purchased from Santa Cruz Biotechnology. HA-Tag (6E2) and Myc-Tag (9B11) antibodies used were purchased from Cell Signaling Technology.

Han B., Copeland C.A., Kawano Y., Rosenzweig E.B., Austin E.D., Shahmirzadi L., Tang S., Raghunathan K., Chung W.K, & Kenworthy A.K. (2016). Characterization of a caveolin-1 mutation associated with both pulmonary arterial hypertension and congenital generalized lipodystrophy. Traffic (Copenhagen, Denmark), 17(12), 1297-1312.

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Western Blot Analysis of Tagged Proteins

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Frozen 6-day-old seedlings were disrupted in a ball mill using metal
beads. Ground tissue was boiled in 2× NuPAGE LDS Sample Buffer (including
1.8% β-mercaptoethanol) for 5 min and separated in NuPAGE 4–12%
Bis–Tris Protein Gels (ThermoFisher Scientific). The following antibodies
were used for immunoblotting: Myc-Tag, 9B11 (1:2,000, catalog no. 2276, Cell
Signaling Technology), anti-HA-peroxidase, high-affinity clone 3F10 (1:2,000,
catalog no. 11867423001, Roche), anti-FLAG M2-peroxidase (HRP) Clone M2
(1:5,000, catalog no. A8592, MilliporeSigma), anti-GFP clones 7.1 and 13.1
(1:5,000, catalog no. 11814460001, Roche), anti-Actin (1:5,000, catalog no.
A0480, MilliporeSigma) and goat anti-mouse IgG (H + L)-HRP conjugate (1:5,000,
catalog no. 1706516, Bio-Rad).

Willige B.C., Zander M., Yoo C.Y., Phan A., Garza R.M., Wanamaker S.A., He Y., Nery J.R., Chen H., Chen M., Ecker J.R, & Chory J. (2021). PHYTOCHROME-INTERACTING FACTORs trigger environmentally responsive chromatin dynamics in plants. Nature genetics, 53(7), 955-961.

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Molecular Toolbox for Cell Signaling

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The following reagents were used in this manuscript; Thapsigargin (T9033/CAY10522, Sigma); Ionomycin (I9657, Sigma); Phorbol 12-myristate 13-acetate (PMA) (79346, Sigma); Fura2-AM, (F1201, Invitrogen); Fluo-8, AM (21082, AAT Bioquest); SiR-Actin (Far Red, Spirochrome) Cyclopiazonic acid from Penicillium cyclopium, (c1530, Sigma); Gadolinium (G7532, Sigma); Vybrant Alexa Fluor 555 Lipid Raft Labeling Kit cholera toxin subunit B (V34404, Thermo-Fisher); Lipophilic Tracer Sampler DiD (L7781, Thermo-Fisher); Hoechst 33,342 (H3570, Thermo-Fisher); Dynabeads Human T-Activator CD3/CD28 (11,161D, Thermo-Fisher), GFP-trap agarose (GTA-10. Chromotek), - hydroxylamine (55460, Sigma, used at 0.5 M), Zebra spin desalting columns (PIER89882, Pierce)), 3H- palmitic acid (ART0129-25, American radio labelled chemicals), NEM (04559, Sigma), protein G (17-0618-01, GEHealthcare), 5 kDa PEG (63187, Sigma). For protein detection either on western blot or immunofluorescence, we used; NFATc1 (clone 7A6, MABS409, Sigma), TCR alpha/beta-PE (12-9986-42, eBioscience), Anti-Cholera Toxin, B-Subunit (227040, Sigma), Myc-Tag (9B11) (2276, Cell-Signalling), gamma Tubulin (4D11) (MA1-850, Thermofisher), ANTI-FLAG M2 (F1804, Sigma), anti-ORAI1 (600–401-DG9, rockland immunochemicals Inc), anti-GFP (SAB4301138, Sigma), anti-mouse-HRP, and rabbit-HRP (1706516 and 172101, Bio-Rad (USA).

Carreras-Sureda A., Abrami L., Ji-Hee K., Wang W.A., Henry C., Frieden M., Didier M., van der Goot F.G, & Demaurex N. (2021). S-acylation by ZDHHC20 targets ORAI1 channels to lipid rafts for efficient Ca2+ signaling by Jurkat T cell receptors at the immune synapse. eLife, 10, e72051.

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Quantification of Autophagy-related Organelles

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Cells were fixed with 4% Pfa, permeabilized with 0.2% Triton, blocked in 1% BSA and were stained with anti-PI(3)P (Echelon), LBPA (Echelon), Atg12 (Cell Signaling #2011), WIPI2 (Abcam), Flag-tag M2 (Sigma), and Myc-tag 9B11 (Cell Signaling) and imaged on upright and inverted confocal microscopes made by Zeiss (Göttingen, Germany) including Zeiss 510 and Zeiss 780. 40× and 63× oil objectives were used. For quantification, at least 30 transfected cells imaged from at least three separate experiments were examined and categorized as indicated (i.e. no rings vs. rings).

McKnight N.C., Zhong Y., Wold M.S., Gong S., Phillips G.R., Dou Z., Zhao Y., Heintz N., Zong W.X, & Yue Z. (2014). Beclin 1 Is Required for Neuron Viability and Regulates Endosome Pathways via the UVRAG-VPS34 Complex. PLoS Genetics, 10(10), e1004626.

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Western Blot Analysis of AMPK Signaling

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Whole cell extracts or 87 HCC tumor specimens were prepared in lysis buffer, and western blot analysis was performed as described previously [50 (link)]. Specific primary antibodies used were as follow: anti-AMPK, phospho-AMPK (Thr172), phospho-ACC (Ser79), and Myc-Tag (9B11) were purchased from Cell Signaling Technology (USA). Antibody against ACCα was obtained from proteintech and β-actin from Santa Cruz Biotechnology (Heidelberg, Germany). After incubating with the fluorescein-conjugated secondary antibody, the immunocomplexes were detected using an Odyssey fluorescence scanner (Li-Cor, Lincoln, NE).

Wang M.D., Wu H., Huang S., Zhang H.L., Qin C.J., Zhao L.H., Fu G.B., Zhou X., Wang X.M., Tang L., Wen W., Yang W., Tang S.H., Cao D., Guo L.N., Zeng M., Wu M.C., Yan H.X, & Wang H.Y. (2016). HBx regulates fatty acid oxidation to promote hepatocellular carcinoma survival during metabolic stress. Oncotarget, 7(6), 6711-6726.

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Antibodies for GlyT2 Protein Analysis

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Male Wistar rats were bred under standard conditions at the Centro de Biología Molecular Severo Ochoa (CBMSO) in accordance with procedures approved in the Directive 2010/63/EU of the European Union with approval of the Research Ethics Committee of the Universidad Autónoma de Madrid (Comité de Ética de la Investigación UAM, CEI-UAM). Antibodies against GlyT2 N-terminus were generated in house (rabbit and rat^{67 (link),68 (link)}) while the other primary antibodies used were: anti-c-Myc (Myc-Tag 9B11 Cell Signaling Tech.), anti-FLAG M2 (Sigma-Aldrich, F3165), anti-HA (Sigma-Aldrich, clon 12CA5), anti-ubiquitin (P4D1, Santa Cruz), anti-α-tubulin (Sigma-Aldrich, T6074). All chemicals used were from Sigma Aldrich unless otherwise noticed. Neurobasal medium and B27 supplement were purchased from Invitrogen.

de la Rocha-Muñoz A., Núñez E., Arribas-González E., López-Corcuera B., Aragón C, & de Juan-Sanz J. (2019). E3 ubiquitin ligases LNX1 and LNX2 are major regulators of the presynaptic glycine transporter GlyT2. Scientific Reports, 9, 14944.

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Western Blotting of Viral Proteins

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Proteins were separated in 4–15% Criterion TGX gels (Bio-Rad, Hercules, CA, USA) and transferred to Trans Blot Turbo Midi Nitrocellulose membranes (Bio-Rad). The membranes were blocked with 5% non-fat dry milk in TBS-T. Antibodies detecting target proteins were diluted in 5% non-fat dry milk in TBS-T and incubated overnight, at 4 °C. Antibodies detecting ORF57 (sc-135746), ORF45 (sc-53883), K8.1 (sc-65446), Actin (sc-8432) were from Santa Cruz Biotechnology (SCBT, Dallas, TX, USA). Antibodies detecting HA tag (HA.11) were from Biolegend (San Diego, CA, USA), Myc-tag (9B11) from Cell Signaling Technology (Danvers, MA, USA) and ORF50 was a gift from Carolina Arias (UC Santa Barbara, Santa Barbara, CA, USA). Anti-Rabbit IgG HRP-linked (7074S) and anti-Mouse IgG HRP-linked (7076S) secondary antibodies were from Cell Signaling Technology. WesternBright Sirius HRP substrate (Advansta, San Jose, CA, USA) was used for revealing the protein bands during imaging in a ChemiDoc gel imaging system (Bio-Rad).

Elbasani E., Falasco F., Gramolelli S., Nurminen V., Günther T., Weltner J., Balboa D., Grundhoff A., Otonkoski T, & Ojala P.M. (2020). Kaposi’s Sarcoma-Associated Herpesvirus Reactivation by Targeting of a dCas9-Based Transcription Activator to the ORF50 Promoter. Viruses, 12(9), 952.

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