Prepare optimized buffers for pull-down and reagent sets of TurboLuc (Tluc) assay buffer and coelenterazine substrate for luciferase quantitation with Thermo Scientific™ Pierce™ qIP Protein Interaction Reagents.
Thermo Scientific Pierce qIP Protein Interaction Reagents include stock solutions to prepare optimized buffers for co-immunoprecipitation (co-IP) and reagent sets of TurboLuc (Tluc) assay buffer and coelenterazine substrate for luciferase quantitation.
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.Highlights:
- 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
- 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.