20 likes | 126 Views
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.<br>
E N D
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