For shorter assay times please try our Immunoprecipitation Protocol Utilizing Magnetic Separation / (For Analysis By Western Immunoblotting).
NOTE: Prepare solutions with Milli-Q or equivalently purified water.
NOTE: Add 1 mM PMSF immediately prior to use.
Optional: It may be necessary to perform a lysate pre-clearing step to reduce non-specific binding to the Protein A/G agarose beads (See section below).
NOTE: For proteins with molecular weights of 50 kDa, we recommend using Mouse Anti-Rabbit IgG (Light-Chain Specific) (L57A3) mAb #3677 or Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb #3678 as a secondary antibody to minimize masking produced by denatured heavy chains. For proteins with molecular weights of 25 kDa, Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb #3678 is recommended.
posted November 2006
revised January 2012