Immunoprecipitation (IP) is a technique to isolate a specific protein out of a solution using an antibody that binds to it. Antibody-protein complexes are removed from the solution with the addition of an insoluble form of an antibody binding protein, such as Protein A or Protein G conjugated to agarose slurry or the newly popular Magnetic Beads. Immunoprecipitation assays detect the interaction of a target protein with other proteins or nucleic acids. Assay examples include Co-IP, ChIP, RIP and tagged protein IP.
Traditional immunoprecipitation (IP) uses agarose beads coated in protein A/G. Most of these IP protocols require more than three washing steps to eliminate background and non-specific binding. Each step of washing causes some bead loss or leaves behind residual contaminants.
The use of magnetic beads has recently gained popularity as a quicker, more accurate approach for immunoprecipitation. Unlike agarose, no columns or centrifugations are required. Also, you can significantly reduce experiment variability by being able to remove all supernatant without disturbing your pellet. You will consistently get more reliable and reproducible data with GenScript MagBeads.
This protocol offers a general guideline for immunoprecipitation with GenScript MagBeads.A. Cross-linking IgG to the Beads
B. Binding Target Protein to the IgG Cross-linked Beads
C. Elution of Target Protein