TrueBlot

Immunoprecipitation (IP) and Western blot (WB) protocols provide highly specific results but often suffer from poor or non-specific binding, high background, and heavy/light chain interference. Rockland’s TrueBlot® line of products consists of both TrueBlot® IP beads and TrueBlot® monoclonal secondary antibodies, which combat these common experimental issues by providing increased specificity, low background, and enhanced accuracy to provide publication-quality IP Western blots without interference from contaminating IgG.

Browse All TrueBlot Products
TrueBlot
TrueBlot Secondary Antibodies
Achieve unparalleled clarity of results and publication-quality images for Western blot and IP-Western blot assays
TrueBlot IP Beads
Optimize and purify complex biological samples faster with TrueBlot IP beads in both agarose and magnetic format
TrueBlot Kits
Harness the unique detection properties of TrueBlot with all the critical reagents combined conveniently in sets and kits

 

TrueBlot_Comparison.png

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.

svg-image

Selecting TrueBlot® Products

Rockland’s TrueBlot® product line allows researchers to generate reproducible, publication-quality IP-Western blots easily. But with so many products to choose from, it may be hard to determine which TrueBlot product is best for your Western blot assay. Use our handy flowchart to navigate to the best TrueBlot product based on your specific assay, detection method, and more.

DOWNLOAD FLOWCHART

Resources

VIEW ALL RESOURCES

TrueBlot® Products

Literature
View Literature

Tips for IP-WB with TrueBlot®

Technical Tips
View Technical Tip

IP-WB with TrueBlot® Protocol

Protocols
View Protocol

When to Use TrueBlot® Products

The TrueBlot® product line is ideal for studying protein-protein interactions, post-translational modifications (phosphorylation and acetylation), poorly expressed proteins, and immunoprecipitated proteins with molecular weights similar to the heavy and light chain.

TrueBlot secondary antibodies preferentially detect the native, non-reduced form of IgG over the reduced form of IgG, making them ideal for Western blots of immunoprecipitated proteins. These unique detection properties reduce interference by the heavy chain (~55 kDa) and light chain (~23 kDa) of the immunoprecipitating antibody enabling unhindered detection of blotted target bands. Pair TrueBlot® antibodies with the exceptional purification properties provided by TrueBlot® IP beads to eliminate the common issues of IP-Western Blot.

TrueBlot® IP beads and TrueBlot® monoclonal secondary antibodies are available in a variety of host species and can be purchased individually, in sets with the corresponding agarose Ig IP beads, or as Western blot kits, allowing researchers the ability to choose a TrueBlot® product to fit all their research needs.