Pierce™ qIP Protein Interaction Assay Reagent Set

Catalog number: 82015

Thermo Scientific™  Related applications: Protein Assays and Analysis

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Description

The Thermo Scientific™ Pierce™ Quantitative Immunoprecipitation (qIP) Protein Interaction Set provides a highly sensitive luciferase-based assay to measure protein-protein interactions in mammalian cells without Western blotting. The qIP assay utilizes the smallest and one of the brightest luciferases, Thermo Scientific™ TurboLuc™ (Tluc) Luciferase, fused to a protein of interest that is transiently co-expressed with an epitope-tagged protein in mammalian cells. The protein interactions are quantified by measuring Tluc Luciferase activity following pull-down of epitope-tagged proteins with anti-epitope agarose or magnetic beads. The Thermo Scientific™ Pierce™ qIP Protein Interaction Tluc Assay Reagents are only optimized for the Tluc Luciferase assay for protein interaction studies.

Features of the qIP protein interaction assay reagents:

Fast and simple – add Buffer L (1X) to mammalian cell pellet and vortex for complete cell lysis
Robust – buffers tested on many different cell types, including but not limited to, HEK293, 293T, NIH3T3, HeLa, CHO-K1, CHO and COS-7 cells for the qIP protein interaction assay
Compatible – completely compatible with different anti-epitope agarose and magnetic beads and TurboLuc (Tluc) luciferase-based qIP assay reagents

Buffer L (supplied at 1X) is the cell lysis buffer developed and optimized to be used with the Pierce qIP Protein Interaction Kits. It is compatible with both agarose resin and magnetic bead formats of these kits. Upon dilution to 1X, Buffer D (supplied at 10X) is used to dilute cell lysates (in Buffer L) to create favorable conditions for Co-IP. The prepared 1X Buffer D is also mixed with Buffer L to make the wash buffer. The Pierce qIP Protein Interaction Tluc Assay Buffer and Coelenterazine Substrate (100X) are used to measure the luciferase activity of protein-protein interactions (Co-IP products) that result from the qIP assay procedure. These reagents are optimized and validated only for Tluc luciferase assays in the Pierce qIP system for protein interaction studies using appropriate expression vectors.

Requires:
• Protease inhibitors (e.g., Halt Protease Inhibitor Cocktail, EDTA-free, Part No. 78437)

The Pierce qIP Protein Interaction System uses anti- HA or anti-Myc beads (agarose or magnetic) and a sensitive luciferase assay system to co-immunoprecipitate (co-IP) and quantify interactions between epitope-tagged and Tluc-tagged protein pairs expressed in mammalian cells. The quantitative immunoprecipitation (qIP) method depends on TurboLuc luciferase enzyme (Tluc) to accurately and precisely reflect the abundance of a specific co-IP product without time-consuming gel electrophoresis, Western blot and band densitometry steps. Complete kits are available with cloning vectors, affinity beads (agarose or magnetic) and assay reagents. Alternatively, all components of the assay system are available separately. This page features the essential buffers and luciferase assay reagents.

More Product Data
Pierce qIP Assay: a novel, HTS-compatible protein-protein interaction assay system

Related Products
Pierce™ qIP Protein Interaction Buffer L (1X)
Pierce™ qIP Protein Interaction Buffer D (10X)
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Technique: Enhanced Chemiluminescence, Immunoprecipitation (IP)
Assay Type: Luciferase Reporter Assay
Product Size: 150 reactions
Detection Method: Bioluminescent
For Use With (Equipment): Luminometer (Microplate)

Contents & storage

Upon receipt, store kit components at -80°C. The Pierce qIP Assay Reagents are shipped on dry ice.

Documents

Manuals & protocols