TrueBlot® Products: Enhanced Sensitivity in IP-Western Blots
Immunoprecipitation (IP) and Western Blot (WB) protocols provide highly specific results, yet can often suffer from poor or non-specific binding, high background, and heavy/light chain interference.
Rockland’s TrueBlot® line of products consist of both TrueBlot® IP beads and TrueBlot® monoclonal secondary antibodies, which combat these common experimental issues by providing increased sensitivity, low background, and enhanced accuracy.
Easy to use – simply substitute your conventional HRP or fluorescent blotting reagent with an appropriate TrueBlot product for unparalleled clarity of results and high-quality images suitable for reproduction and publication.
Features and Benefits
- Detects only the native disulfide(non-reduced) form of IgG
- Reduced interference of heavy and light chain IgG
- Unhindered detection of your blotted protein band of interest.
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.
Get to know more: Watch video the TrueBlot®: Publication Quality IP and Western Blot Images or download our TrueBlot materials
For more information, please contact:
Maria Kam Eriksen
+45 2244 8243