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Protein of interest is obstructed in immunoprecipitation (IP)

Explore potential causes and solutions for protein obstruction in IP.

Protein of interest is obstructed by antibody heavy or light-chains

Possible causes

Solution

Secondary antibody recognizes denatured/reduced primary antibody released during the IP procedure or endogenous IgGs

Using secondary antibodies, recognizing the heavy and light-chain of the primary antibody for WB detection of IP samples will always result in two bands (the heavy-chain at 50kDa and the light-chain at 25kDa).

  • To avoid interference by the antibody chains we recommend using secondary antibodies, such as VeriBlot for IP, that specifically recognize native (non-reduced) primary antibodies.