Background of c-Myc Epitope Tag (EQKLISEEDL) antibody
Epitope tags are short peptide sequences that are easily recognized by tag-specific antibodies. Due to their small size, epitope tags do not affect the tagged protein's biochemical properties. Most often sequences encoding the epitope tag are included with target DNA at the time of cloning to produce fusion proteins containing the epitope tag sequence. This allows anti-epitope tag antibodies to serve as universal detection reagents for any tag containing protein produced by recombinant means. This means that anti-epitope tag antibodies are a useful alternative to generating specific antibodies to identify, immunoprecipitate or immunoaffinity purify a recombinant protein. The anti-epitope tag antibody is usually functional in a variety of antibody-dependent experimental procedures. Expression vectors producing epitope tag fusion proteins are available for a variety of host expression systems including bacteria, yeast, insect and mammalian cells. We offer anti-epitope tag antibodies against many common epitope tags including Myc, GST, GFP, 6X His, MBP, FLAG and HA. Please visit our website at www.acris-antibodies.de for more informations.
General / background references:Spandidos DA et al. Elevated expression of the myc gene in human benign and malignant breast lesions compared to normal tissue. Anticancer Res 7:1299-304 (1987). PubMed PMID: 3327455 Evan GI et al.
Immunolocalization of c-Myc in transfected mammalian cells using SP5385P
Figure 1. Anti-Myc epitope tag polyclonal antibody detects both AMINO and CARBOXY terminal linked Myc-tagged recombinant proteins by Western blot. Polyclonal rabbit-anti-Myc epitope tag antibody was diluted to 1.0 µg/ml to detect either recombinant protein. 4-20% gradient gels were used to separate the proteins by SDS-PAGE. The proteins were transferred to nitrocellulose using standard methods. After blocking the membranes were probed with the primary antibody overnight at 4°C followed by washes and reaction with a 1:10,000 dilution of IRDye® 800 conjugated Gt-a-Rabbit IgG [H&L] for 45 min at room temperature (Green 800 nm channel). Pre-stained molecular weight markers are also shown (lane M, Red 700 nm channel). LICOR's Odyssey® Infrared Imaging System was used to scan and process the image.
MYC-tag polyclonal antibody ( Cat # PAB10345 ) detects ~ 100 kDa carboxy terminal linked MYC-tagged recombinant protein present in ~35 µg of lysate by western blot ( arrowhead ).Carboxy terminal linked MYC recombinant protein was the gift of Zhongsheng You, Salk Institute, LaJolla, CA.
MYC-tag polyclonal antibody ( Cat # PAB10345 ) detects ~60 kDa amino terminal linked MYC-tagged recombinant protein by western blot ( arrowhead ).Lane 1 contains ~35 µg of lysate from control 293T cells.Lane 2 contains ~35 µg of lysate from 293T cells overexpressing an N-terminal linked recombinant protein.Amino terminal linked MYC recombinant protein was the gift of Brian Conti, University of North Carolina, Chapel Hill, NC.
Anti-Myc epitope tag polyclonal antibody detects Myc-tagged recombinant proteins by western blot. Polyclonal rabbit-anti-Myc epitope tag at 0.5-1.0 µg/ml was used to detect 1.0 ug of recombinant protein containing the Myc epitope tag. A 4-20% gradient gel was used to separate the protein by SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking the membrane was probed with the primary antibody for 1 h at room temperature followed by washes and reaction with a 1:2500 dilution of IRDye(TM)800 conjugated Gt-a-Rabbit IgG [H&L] for 30 min at room temperature. LICOR's Odyssey(R) Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results.