Nuclear Protein Immunoprecipitation (IP)

Immunoprecipitation is a technique in which an antigen is isolated by binding to a specific antibody attached to a sedimentable matrix. It is also used to analyze protein fractions separated by other techniques such as gel filtration or density gradient sedimentation. The source of antigen for immunoprecipitation can be unlabeled cells or tissues, metabolically or intrinsically labeled cells, or in vitro-translated proteins. This unit describes a wide range of immunoprecipitation techniques, using either suspension or adherent cells lysed by various means.Usually, plasmas transfected cells or natural cells are ideal sample for immunoprecipitation.

A nuclear protein is a protein found in the cell nucleus. Which binding to other proteins,or DNA or RNA,have important function for the cell's life cycle. If you want to discover the secret of these proteins, you need to extract these proteins from cells or tissues. The protocol of isolate nuclear proteins are viorous,base on your porpose. The buffers used as follow:

Nuclear lysis buffer
10mM Hepes or Tris (pH 7.5)
500mM NaCl
1% Triton-X100
10% glycerol
1mM NaPPi
1ug/mL pepstatin
1ug/mL aprotinin
1ug/mL leupeptin
1mM NaVO4
1mM NaF
water to 50 mL