How TrueBlot® Secondaries Work
When proteins are isolated using IP, a capture antibody binds to the target protein (Step 1), Protein A- or Protein G-coated magnetic beads are added to immobilize the antibody-target protein complex (Step 2) and the undesired material is washed away (Step 3). The final steps in a conventional IP typically include denaturation of the proteins using heat and a reducing/denaturing agent (Step 4).
When the eluted IP sample is separated via SDS-PAGE (Step 5), probed with the same primary antibody used in IP, and detected with conventional secondary antibodies in Western blot, both the heavy and light chain of the IP capture antibody can be detected (Step 5; right panel), which can obscure your protein of interest, specifically if it is similar in size to the heavy or light chain.
TrueBlot® secondary antibodies only recognize primary antibodies in their native (non-reduced) state, eliminating interference from heavy and light chain bands (Step 6; left panel) to provide accurate, publication-quality Western blots of immunoprecipitated samples.