Thermo Scientific™ Pierce™ Direct IP Kit

Thermo Scientific™ Pierce™ Direct IP Kit

The Thermo Scientific Pierce Direct IP Kit uses an activated resin to covalently immobilize IP antibodies (any species or class) on agarose beads without the aid of Protein A/G, enabling immunoprecipitation without antibody interference.

The primary benefits resulting from this method are the opportunity to use any species or subclass of purified antibody (not just types that bind to Protein A or G) and the ability to purify target protein without contamination by the antibody. The method also makes it possible to immunoprecipitate antigens from serum samples without co-purifying non-target immunoglobulins. Finally, the kit uses microcentrifuge spin cups to effectively wash and separate samples from the beaded agarose resin.

Features

• Requires less than 10 µg of antibody—optimized procedure requires only 2 to 10 µg of antibody for a single IP experiment, no more than is required for traditional IP methods
• Easy and efficient scalability—use only the amount of antibody needed for a single immunoprecipitation or immobilize 100 to 200 µg of antibody to prepare ready-made IP affinity resin for many experiments
• Minimal antibody contamination—antibody is irreversibly attached to the agarose beads so that co-elution of heavy and light chains with the purified protein is minimized (i.e., less than 5%)
• IP with any species and subclass of antibody—Use chicken IgY, human IgE, mouse IgM or any other purified protein
• Prepared antibody affinity resin is reusable—because the antibody is covalently immobilized and not inactivated by the mild elution procedure, the resin often can be used several times
• Convenient sample handling—spin columns (see dimensions) eliminate resin loss and provide for efficient separation of solutions
• Complete kit—package includes cell lysis buffer and sufficient reagents and spin columns to perform at least 50 antibody immobilizations and IP experiments
• Requires purified antibody—IP antibody solution must be free of BSA, gelatin or other stabilizer proteins; for easy removal of these proteins, see our Antibody Clean-up Kit (Part No. 44600)

Contents

Preparing and performing at least 50 IP reactions, each with 10 µL of resin

• AminoLink Plus Coupling Resin: 2 mL
• Coupling Buffer (20X): 25 mL
• Quenching Buffer: 50 mL
• Wash Solution: 50 mL
• Sodium Cyanoborohydride Solution (5M): 0.5 mL
• IP Lysis/Wash Buffer: 2 x 50 mL
• Conditioning Buffer (100X): 5 mL
• Tris-Buffered Saline (20X): 25 mL
• Elution Buffer: 50 mL
• Lane Marker Sample Buffer (5X): 5 mL
• Control Agarose Resin: 2 mL
• Spin Columns and Collection Tubes