The Thermo Scientific Pierce c-Myc Tag IP/Co-IP Kit provides the affinity resin and other reagents necessary to easily perform immunoprecipitation (IP) or co-immunoprecipitation (co-IP) experiments using a c-Myc-tagged protein as the bait.
The c-Myc peptide (EQKLISEEDL) has become a popular fusion tag for mammalian recombinant protein expression. This kit includes crosslinked beaded agarose to which a highly specific anti-c-Myc antibody is covalently immobilized. Upon incubation with a sample containing the tagged fusion protein, interaction complexes involving the c-Myc-tagged bait protein are captured on the agarose beads. After simple washing steps, the specific protein interaction complex is easily eluted from the resin in the supplied elution buffer or SDS-PAGE sample loading buffer for subsequent analysis. The kit includes the prepared agarose affinity resin, buffers, microcentrifuge spin columns, a positive control and easy-to-follow instructions.
- Specific – the immobilized anti-c-Myc monoclonal antibody binds the c-Myc epitope tag with high specificity, providing high yield immunoprecipitation products and clean Western blot detection
- High capacity – excellent results with as little as 1µg of anti-c-Myc antibody in IP mode with the positive control lysate
- Rigorous – kit includes a positive control lysate that contains over-expressed GST-c-Myc to validate the reagents and specific protocol, and the instructions provide tips for planning other controls
- Robust – the affinity system is compatible with IP or Co-IP from various cell lysates and physiological (non-denaturing) buffer systems
- Convenient and easy – complete kit includes all necessary reagents, convenient spin columns, and easy-to-follow instructions
The c-Myc peptide (EQKLISEEDL) derived from the C-terminus region of human c-Myc protein is one of several fusion protein tags used for recombinant protein expression. Utilizing a specific, high-affinity immobilized antibody, c-Myc tagged fusion proteins can be quickly purified from bacterial and mammalian cell lysates as well as from the Pierce Human in vitro translation reactions. For co-immunoprecipitation reactions, simple wash steps allow enrichment and elution of specific protein interaction complexes into the supplied elution buffer or SDS-PAGE sample loading buffer for subsequent analysis.
|High-quality immunoprecipitation of c-Myc-tagged protein. Benchmark comparison of two anti-c-Myc agarose resins for small-scale purification (IP) performance. Pierce Anti-c-Myc Agarose provides cleaner (more specific) purification and equal yield.
Co-IP of BCL2-Luc with c-Myc-tagged BAD in 293T cells. This experiment tests the specificity of co-immunoprecipitation by comparing results from 293T cells cultured alone (293T) or with vectors expressing BCL2-Luc and/or BAD-c-Myc fusion proteins (labeled BCL2 and BAD, respectively). The Western blot reveals the presence of BCL2 detected through its fused luciferase domain (i.e., via an anti-Luc antibody). Although BCL2 is expressed in two cultures (as indicated by its detection in two of the Control Lysates), it is purified by anti-c-Myc agarose beads (IP or Co-IP lanes) only when co-expressed and captured through its interaction with c-Myc-tagged BAD (BAD+BCL2 lane).
Epitope Tag Product Selection Guide – links to antibodies, resins, magnetic beads and IP kits
IgG Elution Buffer
Other Anti-Tag Antibodies
Fusion Protein Purification Resins and Kits
Cell Lysis and Protein Extraction Reagents