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Immunoprecipitation (IP) is a method of precipitating target protein out of solution by using an antibody that specifically binds to that particular protein. This process is powerful to isolate and concentrate a particular protein and/or protein complex from a sample containing many thousands of different proteins.\n

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immunoprecipitation guide

Immunoprecipitation Guide

Immunoprecipitation (IP) is a method of precipitating target protein out of solution

by using an antibody that specifically binds to that particular protein. This process is

powerful to isolate and concentrate a particular protein and/or protein complex from

a sample containing many thousands of different proteins.

Basic procedure

The basic procedure of IP or Co-IP begins with the extraction of antigen from tissue

samples or cells in an appropriate lysis buffer. Particularly for ChIP or RIP, the nucleic

acid is cross-linked to protein by using formaldehyde or UV irradiation before cell

lysis. The resulting lysate is incubated with an antibody, allowing the formation of an

immune complex. Immobilized Protein A or Protein G linked to agarose or magnetic

beads is simultaneously added to provide a solid phase support, which enables the

precipitation

or

capture

of

those

immunoprecipitated complexes are collected by centrifugation, and proteins are

eluted for further analysis such as western blot and LC/MS. Nucleic acid analysis is

achieved through PCR, RT-PCR, cDNA sequencing and microarray.

antibody-antigen

complexes.

The

https://www.creative-diagnostics.com/immunoprecipitation-guide.htm