Conjugate any antibody or protein to magnetic beads and immunoprecipitate (affinity purify) target protein complexes without IP-antibody interference.
Brand: Thermo Scientific™ 88828
556.80 EUR valid until 2025-06-30
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Thermo Scientific Pierce Direct Magnetic IP/Co-IP Kit uses NHS-chemistry to covalently immobilize IP antibodies onto magnetic beads for effective immunoprecipitation and co-immunoprecipitation.
Magnetic IP with NHS chemistry is convenient and allows for the immobilization of IP antibody from any species and subtype without the variations in coupling efficiency seen with Protein A and Protein G. The IP antibody is covalently attached so target proteins or co-IP protein complexes can be eluted and analyzed without antibody contamination. The kit contains an optimized protocol and all buffers and reagents necessary for antibody coupling and high yield IP or co-IP using either manual magnetic stands or automated platforms such as the Thermo Scientific KingFisher Instruments.
Features of the Direct Magnetic IP/Co-IP Kit:
The protocol for the Pierce Direct IP/Co-IP Kit conjugates IP antibody to Pierce NHS-Activated Magnetic Beads (Catalog #88826) through NHS-ester chemistry. The antibody-bound beads are then incubated with quenching buffer to block remaining active NHS sites. The prepared antibody-bound beads are incubated with cell lysate or tissue extract that contain the protein antigen of interest, allowing the antigen:antibody complex to form. The beads are washed to remove nonbound material and a low-pH elution buffer is used to dissociate bound antigen from the antibody-bound bead. Neutralization buffer is included to prevent precipitation of the isolated antigen and to ensure protein activity in downstream applications. Lane Marker Sample Buffer is included for preparing samples for SDS-PAGE analysis. The protocol is optimized for 2 to 10ÎĽg of IP antibody per assay. For optimal co-IP yields, use 10ÎĽg of antibody. The beads can be processed manually using a magnetic stand or by automation with an instrument such as the Thermo Scientific KingFisher Flex.
Traditional IP with Protein A/G Magnetic Beads (Catalog #88804), involving passive, noncovalent coupling of antibody with Protein A/G typically provides higher antigen yields than IP protocols that involve covalent attachment of antibody to magnetic beads through crosslinking or NHS-ester chemistry. Covalent attachment inevitably causes loss of some functional antibody binding sites. To overcome this limitation with NHS-chemistry, it may be necessary to use larger amounts of IP antibody in the direct IP method than in the traditional IP method. The advantage of immunoprecipitation with IP antibody covalently bound to the magnetic bead is Western blots and gels devoid of interfering antibody bands.
Related Products
Pierce™ Direct IP Kit (Catalog #26148)
| 1 mL | |
| Sufficient For: 40 IP reactions using 25 μL of beads • Pierce NHS-Activated Magnetic Beads, 1 mL • IP Lysis/Wash Buffer, 2 x 50 mL • Elution buffer, pH 2.0, 5 mL • Lane Marker Sample Buffer, Non-reducing, (5X), 5 mL • Neutralization Buffer, pH 8.5, 0.5 mL • 0.67M Borate Buffer, 1 mL • BupH Borate Buffer Pack, 1 pack • Quenching Buffer, 25 mL Store at 4°C. |
|
| KingFisher™ Flex, Magnetic Stand | |
| 40 reactions | |
| Not Target-Specific | |
| 40 Reactions |
| Slurry: 10 mg/mL, 1% solids | |
| Affinity Purification | |
| Pierce™ | |
| Pull-Down, Immunoprecipitation (IP) | |
| Immunoprecipitation/Co-Immunoprecipitation Kit | |
| Kit |
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Catalog No. 17767453
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Expires: 2025-06-27
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For Research Use Only. Not for use in diagnostic procedures.
