Thermo Scientific Pierce Classic Magnetic IP/Co-IP Kit uses less than 10 μg of antibody and a magnetic separator. Highly effective and efficient immunoprecipitation (IP) and co-immunoprecipitation (co-IP) of antigens and protein complexes.

Features of the classic magnetic bead method IP/Co-IP kit:

•Compatible—Magnetic beads based on protein A/G recombinant proteins ensure compatibility with large Compatible with most primary antibodies (whether from mouse or rabbit)
• Fast—immunoprecipitation is as short as 30 minutes to reduce background and increase capture of transient protein complexes
• < b>Clean—Immobilize your antibodies to prevent contamination of the eluate
• Tolerant—In the presence of detergents, low pH buffers, or common mass spectrometry solvents No leaching of Protein A/G
• Efficient—uses half the recommended volume of magnetic particles for immunoprecipitation compared to other magnetic beads
• Convenience< /b>—Comprehensive Co-IP kit contains magnetic beads and all necessary buffers required for a complete immunoprecipitation experiment

This immunoprecipitation kit uses high-quality Thermo Scientific Pierce Protein A/G magnetic beads and optimization Buffers and flexible protocols for high-yield IP or co-IP using manual or automated magnetic separation tools. Optimized lysis/wash buffer optimizes yield and efficient antibody-antigen binding and co-IP interactions. A relatively mild, low pH elution buffer separates bound immune complexes from Protein A/G. Alternatively, the included lane marker sample buffer provides fast 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 Loading Buffer
< br>Immunoprecipitation using magnetic particles is very similar to beaded agarose IP, except that a magnet is used instead of centrifugation for separation. Specific antibodies are first added to the sample to form an immune complex, which is then bound to the magnetic beads. The complexes are washed to remove unbound material, and a low pH elution buffer separates bound immune complexes from Protein A/G. Alternatively, include lane marker sample buffer for dissociation using denaturing conditions or for downstream sample preparation for SDS-PAGE analysis. This kit contains Thermo Scientific Pierce Protein A/G Magnetic Beads (for quick and easy detection of antigens magnetic separation) and optimized buffers (for improved antigen yield). Magnetic beads are removed from solution manually using a magnetic stand or through automation using an instrument such as the Thermo Scientific KingFisher Flex instrument.

It is generally known that longer antibody-sample incubations (2 hours to overnight) generally provide higher yields in IP assays. It is often thought that this higher yield is related to increased background. Our researchers have found that overnight antibody-sample binding followed by immunoprecipitation using the Pierce Classic Magnetic IP/Co-IP Kit generally improves yields without increasing background. Therefore, we recommend that researchers perform this binding step within at least 1 hour if possible.

Protocol Summary:
• Incubate cell lysates with IP antibodies for 1 to 2 hours at room temperature or overnight at 4ºC.
• Allow antigen-antibody complexes to bind to Protein A/G magnetic beads for 1 hour at room temperature.
•Wash the beads twice with IP lysis/wash buffer and once with purified water.
•Elute antigen/antibody complexes.

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