ChromoBlog

Co-immunoprecipitation (Co-IP) describes the isolation of a protein and its binding partners from a cell extract using a Nanobody or antibody that is bound to beads. The protein, that directly interacts with the Nanobody, or antibody beads is called “bait”. The binding partner that is indirectly precipitated is called “prey”.

Co-IP describes the immunoprecipitation of a protein of interest and its interacting partners.

When time in the lab is limited, learn how to save precious hours on your immunoprecipitation, immunofluorescence and western blotting experiments.

Due to the current situation with the COVID-19 pandemic, many researchers around the world are faced with restrictions on time and resources in the lab. As we adapt to these changes, the pressure is on to find ways to save time without compromising on results. Proteintech and ChromoTek offer solutions to save you hours on your current immunoassay protocols, helping you achieve publication worthy results in a fast and reliable manner.

V5 tag is a short peptide tag for detection and purification of proteins. The V5 tag can be fused/cloned to a recombinant protein and detected in ELISA, flow cytometry, immunoprecipitation, immunofluorescence, and Western blotting with antibodies and Nanobodies.

ChromoTek's Nano-Traps are optimized for the immunoprecipitation (IP) of proteins and their interacting factors. Nano-Traps comprise of a Nanobody/ V_HH conjugated to beads. For 3 Nano-Traps, e.g. GFP-Trap^®,  RFP-Trap^®, and Spot-Trap^®, we offer different types of beads:

• Agarose beads
• Magnetic Agarose beads
• Magnetic Particles M-270

Immunoprecipitation with GFP-Trap.
I: Input, FT: Flow-Through, B: Bound

Based on the properties of the different beads (see table), we recommend:

• Agarose beads for very low background and high binding capacity IP
• Magnetic Agarose beads for magnetic separation and high binding capacity IP
• Magnetic Particles M-270 for IP of very large proteins/complexes
  

Introduction of Affinity vs. Specificity

Introduction

A high background from unspecific binding of proteins is a common problem in immunoprecipitation (IP). In this blog, the origin of background and different optimization strategies are discussed. Furthermore, it is shown how you can achieve a low background when using the ChromoTek GFP-Trap^®, a ready to use reagent for IP of a GFP-tagged proteins.

Estimated reading time:  3 minutes

The pull-down of proteins is an established technology. It can be difficult, particularly when the protein of interest is expressed at low levels. Here we discuss 5 tips that help you to improve your IP results.

Affinity tags are valuable tools for immunoprecipitation (IP). Fused to the protein of interest, they enable proteome-wide identification of interaction partners by mass spectrometry.