1. Dr. P. Saranraj
Head
Department of Microbiology
Sacred Heart College (Autonomous)
Tirupattur – 635 601
Tamil Nadu, India
Mobile: +91-9994146964; E.mail:
microsaranraj@gmail.com
IMMUNOPRECIPITATION
2. IMMUNOPRECIPITATION
Immunoprecipitation (IP) is a method to isolate a
specific antigen from a mixture, using the Ag -
Ab interaction.
Antigens isolated by IP are analyzed by SDS-
PAGE or Western blotting.
In IP, an antibody is added first to a mixture
containing an antigen, and incubated overnight
at 4 °C under Shaking condition to allow Ag - Ab
complexes to form.
Subsequently, the antigen-antibody complexes
are incubated overnight at 4 °C under Shaking
condition with an immobilized antibody against
the primary antibody (secondary antibody) or
with Protein A/G-coated beads to allow them to
3.
4. Applications of Immunoprecipitation
Isolate / Detect Proteins of interest
Target gene identification.
Enrichment of low abundant proteins.
Study protein-protein interaction and protein
complexes.
Identify unknown proteins in a protein complex
Verify protein expression in a specific tissue.
Determining Size/Molecular weight of protein.
Determining Specific enzyme activity.