Myc Tag antibody (Agarose)

Antigen: Myc Tag

Clonality: Monoclonal (PL14)

Host: Mouse

Conjugate: Agarose Beads

Application: Immunoprecipitation (IP)

Catalog no.: ABIN131876

Additional Information

Immunogen: This antibody was purified from hybridoma (clone PL14) supernatant using protein A agarose. This hybridoma was established by fusion of mouse myeloma cell P3U1 with Balb/c mouse splenocyte immunized with GST-6myc-Tag fusion protein.

Isotype: IgG1

Clone: PL14

Description: Epitope tagging is a widely accepted technique that fuses an epitope-containing peptide to a target protein as a marker for gene expression. With this technique, the gene expression can be easily monitored on western blotting, immunoprecipitation and immunofluorescence utilizing with an antibody that recognizes such an epitope. Amino acid sequences that are widely used for the epitope tagging are as follow, YPYDVPDYA (HA-Tag), EQKLISEEDL (Myc-Tag) and YTDIEMNRLGK (VAV-G- Tag), which corresponding to the partial peptide of Influenza hemagglutinin protein, Human c-myc gene product and Vesicular stomatitis virus glycoprotein respectively.

Specificity: This antibody recognizes Myc-Tag peptide sequence (EQKLISEEDL) on Immunoprecipitation.

Application Details

Application Notes: Immunoprecipitation 1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50 mM Tris-HCl pH 7.2, 250 mM NaCl, 0.1% NP-40, 2 mM EDTA, 10% glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds) 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. 3) Add 20 µL (a 50% gel slurry) of Agarose conjugated Anti-Myc Tag monoclonal antibody into 200 µL of the supernatant. Mix well and incubate with gentle agitation for 30-120 minutes at 4 o C. 4) Wash the agarose 3-5 times with the cold Lysis buffer (centrifuge the tube at 2,500 x g for 10 seconds). 5) Resuspend the agarose in 20 µL of Laemmli’s sample buffer, boil for 3-5 minutes, and centrifuge for 5 minutes. 6) SDS-PAGE & Western Blotting analysis can be performed on a sample of the supernatant. 060609-1 Immunoprecipitation: 20 µL / 200 µL of cell extract from 5x10 6 cells

Buffer: 200 µg of anti-Myc Tag monoclonal antibody covalently coupled to 200 µL of agarose gel and provided as a 50% gel slurry suspended in PBS containing preservative (0.09% sodium azide) for a total volume of 400 µL. *Azide may react with copper or lead in plu

Research Area: Tags/Labels

Restrictions: For Research Use only

Images

Immunoprecipitation of CgA-Myc fusion protein from culture supernatant of CgA-Myc fusion protein expressed 293T cells with ABIN131876 (1), M047-3 (2) or mouse IgG1 (3). After immunoprecipitated with the antibody, immunocomplex was resolved on SDS-PAGE and immunoblotted with M047-3

Publications

Publications: Derijard, Raingeaud, Barrett et al.: "Independent human MAP-kinase signal transduction pathways defined by MEK and MKK isoforms." in: Science (New York, N.Y.), Vol. 267, Issue 5198, pp. 682-5, 1995 (PubMed).

Bergerat, de Massy, Gadelle et al.: "An atypical topoisomerase II from Archaea with implications for meiotic recombination." in: Nature, Vol. 386, Issue 6623, pp. 414-7, 1997 (PubMed).

Bergamaschini, Canziani, Bottasso et al.: "Alzheimer's beta-amyloid peptides can activate the early components of complement classical pathway in a C1q-independent manner." in: Clinical and experimental immunology, Vol. 115, Issue 3, pp. 526-33, 1999 (PubMed).

Mukai, Hachiya, Shoji-Hoshino et al.: "NADE, a p75NTR-associated cell death executor, is involved in signal transduction mediated by the common neurotrophin receptor p75NTR." in: The Journal of biological chemistry, Vol. 275, Issue 23, pp. 17566-70, 2000 (PubMed).

Fukumoto, Watanabe-Fukunaga, Fujisawa et al.: "The fused protein kinase regulates Hedgehog-stimulated transcriptional activation in Drosophila Schneider 2 cells." in: The Journal of biological chemistry, Vol. 276, Issue 42, pp. 38441-8, 2001 (PubMed).

Sasaki, Masuda, Iwai et al.: "A RING finger protein Praja1 regulates Dlx5-dependent transcription through its ubiquitin ligase activity for the Dlx/Msx-interacting MAGE/Necdin family protein, Dlxin-1." in: The Journal of biological chemistry, Vol. 277, Issue 25, pp. 22541-6, 2002 (PubMed).

Ohya, Kawasaki, Hiraga et al.: "The DNA polymerase domain of pol(epsilon) is required for rapid, efficient, and highly accurate chromosomal DNA replication, telomere length maintenance, and normal cell senescence in Saccharomyces cerevisiae." in: The Journal of biological chemistry, Vol. 277, Issue 31, pp. 28099-108, 2002 (PubMed).

Tojo, Matsuzaki, Minami et al.: "The aryl hydrocarbon receptor nuclear transporter is modulated by the SUMO-1 conjugation system." in: The Journal of biological chemistry, Vol. 277, Issue 48, pp. 46576-85, 2002 (PubMed).

Hirota, Daitoku, Matsuzaki et al.: "Hepatocyte nuclear factor-4 is a novel downstream target of insulin via FKHR as a signal-regulated transcriptional inhibitor." in: The Journal of biological chemistry, Vol. 278, Issue 15, pp. 13056-60, 2003 (PubMed).