Brief Presentation on Immuno and Coimmuno precipitation

1. C0-IMMUNO
PRECIPITATION

2. IMMUNO-PRECIPITATION
• Immunoprecipitation is a precipitation technique that uses an
antibody to isolate the antigen of interest from a solution.
• The antibody binds to the antigen and the antibody/antigen
complex is pulled out of the sample using protein A/G-coupled
agarose or magnetic beads.
• The beads are washed and the antigen is eluted.
• Purified antigen obtained by IP is verified by a variety of
molecular techniques (e.g., ELISA and Western blot).

4. CO-IMMUNO
PRECIPITATION
 Co-Immunoprecipitation (Co-IP) is a powerful tool used to
analyze protein-protein interactions.
 The main purpose of Co-IP is the identification of interaction
partners (other proteins) to the protein of interest.
 It is an effective process used to separate proteins from serum,
cell lysate, homogenized tissue, or conditioned media.

5. CO-IMMUNO PRECIPITATION
 Coimmunoprecipitation generally works by preparing a specific antibody that targets a
known protein (antigen) which is believed to be a member of a larger complex of
proteins.
 When the antibody is added to the sample, it binds to a specific protein in the protein
complex.
 Then agarose or magnetic beads are coupled with protein A/G.
 The beads bind to the Antibody bound to the protein complex through the Fc portion.
 After centrifugation the antibody along with the protein complex is precipitated.

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7. CONT…
 Then it is run on SDS-PAGE to separate proteins on
basis of size.
 To see whether the expected protein is present along with
the protein of interest, western blotting is done.
 For the identification of proteins that actually which
protein is present along with proteins of interest, is done
through Mass spectroscopy.