The Thermo Scientific Pierce Classic Magnetic IP/Co-IP Kit enables highly effective and efficient immunoprecipitation (IP) and co-immunoprecipitation (co-IP) of antigens and protein-complexes using less than 10 µg of antibody and a magnetic separator.
Features of the Classic Magnetic IP/Co-IP Kit:
• Compatible—magnetic beads based on Protein A/G recombinant protein ensures compatibility with most primary antibodies, whether from mouse or rabbit • Fast—immunoprecipitate in as few as 30 minutes to reduce background and improve the capture of transient protein complexes • Clean—immobilize your antibody to prevent contamination in your eluate • Resistant—no leaching of Protein A/G in the presence of detergents, low pH buffers or common mass spectrometry solvents • Efficient—immunoprecipitate with half the recommended volume of magnetic particles compared to other magnetic beads • Convenient—the comprehensive Co-IP kit contains the magnetic beads and all essential buffers to complete immunoprecipitation experiments
This immunoprecipitation kit uses high-quality Thermo Scientific Pierce Protein A/G Magnetic Beads and optimized buffers and flexible protocol to accomplish high-yield IP or co-IP with manual or automated magnetic-separation tools. The optimized Lysis/Wash Buffer optimizes yield and efficient binding of antibody-antigen and co-IP interactions. The relatively gentle, low-pH Elution Buffer dissociates the bound immune complex from the Protein A/G. Alternatively, the included Lane Marker Sample Buffer provides rapid, denaturing elution for direct SDS-PAGE analysis of IP products.
Includes: Pierce Protein A/G Magnetic Beads, lysis/wash buffer, elution buffer, neutralization buffer and sample loading buffer
Immunoprecipitation with magnetic particles is performed much like IP with beaded agarose, except that separations are performed using a magnet rather than by centrifugation. The specific antibody is first added to the sample to form an immune complex that is then bound to the magnetic beads. The complex is washed to remove non-bound material and a low-pH elution buffer dissociates the bound immune complex from the Protein A/G. Alternatively, the Lane Marker Sample Buffer is included for dissociation using denaturing conditions or for downstream sample prep for SDS-PAGE analysis. The kit includes Thermo Scientific Pierce Protein A/G Magnetic Beads for fast and convenient magnetic isolation of antigens and optimized buffers for high antigen yield. The beads are removed from the solution manually using a magnetic stand or by automation with an instrument such as the Thermo Scientific KingFisher Flex Instrument.
It is generally known that longer antibody-sample incubations times (2 hours to overnight) usually provide greater yields in IP assays. It is often assumed that this greater yield comes with increased background. Our researchers have found that overnight antibody-sample binding followed by immunoprecipitation with the Pierce Classic Magnetic IP/Co-IP Kit does tend to increase yield but does not increase background. Thus, we recommend that researchers perform this binding step for at least 1 hour whenever possible.
Protocol Summary: • Incubate cell lysate with IP antibody for 1 to 2 hours at room temperature or overnight at 4ºC. • Bind antigen-antibody complex to Protein A/G magnetic beads for 1 hour at room temperature. • Wash beads twice with IP Lysis/Wash Buffer and once with purified water. • Elute the antigen/antibody complex.
Sufficient For: 40 IP reactions using 25 µL of beads • Pierce Protein A/G Magnetic Beads, 1 mL • Pierce IP Lysis/Wash Buffer, 2 x 50 mL • Lane Marker Sample Buffer (5X), 5 mL • Elution Buffer, 5 mL • Neutralization Buffer, 0.5 mL