Overview: Immunoprecipitation (IP) is a method that uses the antigen-antibody reaction principle to identify a protein that reacts specifically with an antibody from mixture of proteins so that its quantity or physical characteristics can be examined. A typical immunoprecipitation experiment contains the following steps. 1) The proteins from the cell or tissue homogenate are precipitated in an appropriate lysis buffer containing an immune complex which includes the antigen (protein), primary antibody and Protein A-, G-, or L-agarose conjugate (Protein A or G binds to the antibody, which is bound to its antigen) or a secondary antibody-agarose conjugate. 2) The immune complex is then captured on a solid support to which either Protein A or Protein G has been immobilized. Proteins that bind to the antibody are precipitated and proteins that do not are washed away. 3) Components of the bound immune complex (both antigen and antibody) are eluted from the support. 4) Immunoprecipited proteins are further analyzed by SDS-PAGE and immunoblotting to determine their molecular weights, to study their interactions with other proteins, to determine specific enzymatic activity, to monitor protein post-translational modifications and to determine the presence and quantity of proteins.
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Protocols
- Immunoprecipitation of Proteins (Upstate)
Well written protocol from Upstate for general use.
http://www.millipore.com/immunodetection/id3/immun...
Added: Tue Feb 24 2004, Hits: 42820, Reviews: 0 Write review Cached - Immunoprecipitation (IP) Protocols (Sigma)
Describes two methods of immunoprecipitation of specific antigens and preparation for SDS-PAGE,
http://www.sigmaaldrich.com/Area_of_Interest/Life_...
Added: Sat Mar 17 2007, Hits: 7613, Reviews: 0 Write review Cached - Immunoprecipitation Protocol (eBioscience)
Immunoprecipitation involves the interaction between a protein and its specific antibody, the separation of these immune complexes with Protein G or Protein A, and the subsequent analysis by SDS-PAGE. This technique provides a rapid and simple means to separate a specific protein from whole cell lysates or culture supernatants. This protocol offers a general guideline for immunoprecipitation.
http://www.ebioscience.com/ebioscience/appls/IP.ht...
Added: Mon Jan 02 2006, Hits: 10122, Reviews: 0 Write review Cached - Metabolic Labeling & Immunoprecipitation (Springer Lab, Harvard University)
Includes radiolabeling and immunoprecipitation in great details.
http://www.cbrinstitute.org/investigators/springer...
Added: Tue Aug 13 2002, Hits: 7552, Reviews: 0 Write review Cached -
Analysis of Protein by Immunoprecipitatioin
(P.J. Hansen Lab)
This protocol describes two methods (protein A and G) in great details as well as the principal, application and trouble shooting of immunoprecipitation.
http://www.dps.ufl.edu/hansen/protocols/imp98.prt....
Added: Tue May 14 2002, Hits: 20993, Reviews: 1 Read reviews Write review Cached - Antibody Precipitation with Protein A or G (Contributed by James Turmo)
Using Immobilized Protein A or Protein G for Immunoprecipitation.
Added: Tue May 14 2002, Reviews: 0 Write review -
Cold Immunoprecipitations
(Bjorkman Group, Howard Hughes Medical Institute at California Institute of Technology)
For screening soluble FCRN mutant transfected cell supernatants
http://www.its.caltech.edu/%7Ebjorker/Immunopptns_...
Added: Wed Aug 14 2002, Hits: 4877, Reviews: 0 Write review -
Cytosolic Extract & Immunoprecipitation for C. elegans
(Schedl Lab, Washington University School of Medicine)
http://www.genetics.wustl.edu/tslab/Protocols/Cyto...
Added: Wed Jul 26 2006, Hits: 1060, Reviews: 0 Write review Cached -
General Principles of Immunoprecipitation
(Sefton Lab, Salk Institute)
http://pingu.salk.edu/~sefton/Hyper_protocols/immu...
Added: Tue Aug 13 2002, Hits: 17440, Reviews: 0 Write review Cached - Glycolipid Binding Assay (Contributed by Pingsunjim, Paller’s Lab)
This protocol can be used for the detection of glycolipids binding to immunoprecipited protein.
Added: Tue May 14 2002, Reviews: 0 Write review