Pierce™ Classic IP Kit

Catalog number: 26146

Thermo Scientific™  Related applications: Protein Assays and Analysis

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Description

The Thermo Scientific Pierce Classic IP Kit uses high-quality Protein A/G agarose beads, microcentrifuge spin columns and buffers to perform traditional immunoprecipitation assays with ease.

The kit uses high-capacity Protein A/G agarose affinity resin for efficient binding of most species and subclasses of IP antibodies. The included IgG elution buffer provides milder and less denaturing recovery of antibody:antigen complexes than the traditional method of boiling in reducing sample buffer for SDS-PAGE, facilitating a greater variety of methods for subsequent analysis. The microcentrifuge spin column format helps to ensure effective washing and separation of samples from the beaded agarose affinity resin.

Features of the Classic IP Kit:

Assay consistency—spin columns (see dimensions) eliminate resin loss and provide for more efficient separation of solutions than traditional IP methods that use only microcentrifuge tubes
Fast—immunoprecipitate (IP) in less than one hour
Mild elution conditions—recover antigen without harsh detergents or reducing agents
Easy-to-follow instructions—purify target protein from crude lysate in four simple steps
Suitable for most common antibody types—Protein A/G provides excellent binding for most mouse, rabbit, human and goat IgG subclasses
Complete kit—includes lysis/wash buffer, binding and elution buffers, control agarose resin, sample loading buffer and spin columns

Applications:
• Routine, traditional immunoprecipitation experiments
• Immunoprecipitation for analysis in nonreducing conditions

Like traditional IP methods, the Pierce Classic IP Kit procedure involves formation of antibody:antigen complexes in a sample solution and then capture of that complex to an IgG-binding protein that is covalently bound to beaded agarose resin (Protein A/G Agarose). After washing to remove nonbound (presumably undesired) components of the sample, the antigen and antibody are recovered from the beaded resin with elution buffer supplied in the kit. The entire procedure is performed in a microcentrifuge spin column, allowing solutions to be fully separated from the agarose resin upon brief centrifugation.

Related Products
Pierce™ Classic Magnetic IP/Co-IP Kit
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Technique: Immunoprecipitation (IP)
Assay Type: Affinity Purification
Product Size: 50 reactions
Target Specificity: Not Target-Specific
For Use With (Equipment): Microcentrifuge

Contents & storage

Sufficient For: At least 50 IP reactions with 10 µL resin
• Protein A/G Plus Agarose, 0.55 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

Store at 4°C.

Documents

Manuals & protocols