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Thermo Scientific™ Pierce™ Classic IP Kit

Catalog No. PI26146
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50 reactions kit
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PI26146 50 reactions kit
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Catalog No. PI26146 Supplier Thermo Scientific™ Supplier No. LSG26146
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Description

Perform traditional immunoprecipitation assays with ease using high-quality Protein A/G agarose beads, microcentrifuge spin columns and buffers.

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

The Classic IP Kit provides all the necessary reagents, spin cups and collection tubes to easily perform successful immunoprecipitation (IP) experiments. 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.

Like traditional IP methods, the Pierce Classic IP protocol 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.

Highlights:

  • Assay consistency – spin columns eliminate resin loss, improve sample recovery and ensure efficient separation from resin

  • Easy protocol – immunoprecipitate (IP) target proteins from crude lysates in just four easy steps

  • Fast – complete an entire immunoprecipitation experiment in less than one hour

  • Economical – reuse the Protein A/G agarose resin for future immunoprecipitations

  • Complete – kit includes a compatible cell lysis reagent and sufficient components to perform at least 50 IP experiments

Includes:

Pierce Protein A/G Plus Agarose, lysis/wash buffer, conditioning buffer, Tris-buffered saline, elution buffer, lane marker sample buffer, control agarose resin, spin columns and collection tubes

Recommended for:

Routine, traditional immunoprecipitation experiments; Immunoprecipitation for analysis in nonreducing conditions

Specifications

Content And 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.
Assay Affinity Purification
For Use With (Equipment) Microcentrifuge
Product Line Pierce
Quantity 50 reactions kit
For Use With (Application) Immunoprecipitation (IP)
Target Not Target-Specific
Type Immunoprecipitation/Co-Immunoprecipitation Kit
Sufficient For 50 Reactions
Format Kit

Frequently Asked Questions (FAQs)

For the Pierce Classic IP Kit protocol, Step B says to add 80 µL of Control Agarose Resin slurry into a spin column for 1 mg lysate. Does the 1 mg of lysate refer to the amount of protein in the lysate?

The "1 mg of lysate" mentioned in Step B of the user manual refers to the total amount of protein in the lysate. The binding capacity of the Control Agarose Resin slurry is 1 mg of lysate (protein) per 80 µl (40 µl of settled resin) of resin slurry.

For Research Use Only. Not for use in diagnostic procedures.