c-Myc (MYC) Mouse Monoclonal Antibody [Clone ID: PL14]

CAT#: AM26688AG-N

c-Myc (MYC) mouse monoclonal antibody, clone PL14, Agarose

Product Images

Immunoprecipitation of CgA-Myc fusion protein from culture supernatant of CgA-Myc fusion protein expressed 293T cells with AM26688AG-N (1), AM26688AF-N (2) or mouse IgG1 (3). After immunoprecipitated with the antibody, immunocomplex was resolved on SDS-PAGE and immunoblotted with AM26688AF-N

Specifications

Product Data

• Clone Name: PL14
• Applications: IP
• Recommended Dilution: Immunoprecipitation: 20 μl / 200 μl of cell extract from 5x10⁶ cells. For details see protocol below.
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: GST-6myc-Tag fusion protein
• Specificity:

  This antibody recognizes Myc-Tag peptide sequence (EQKLISEEDL).

• Formulation: 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
  Label: Agarose
  State: Liquid Ig fraction
• Concentration: lot specific
• Purification: Protein A agarose
• Conjugation: Agarose
• Storage:

  Store at 2 - 8 °C.

• Stability: Shelf life: one year from despatch.

• Gene Name: v-myc avian myelocytomatosis viral oncogene homolog
• Database Link:
  UniProt ID P01106
• Background: 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.
• Synonyms: myc tag, myc-tag, c-myc tag
• Note: This product was originally produced by MBL International.
  
  Protocol:

  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 oC 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 oC 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 oC.
  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.