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Blocking buffers are used in various applications to decrease non-specific signaling generated by non-specific binding of proteins or peptides, essentially blocking western blot noise. Assay sensitivity improves by utilizing the appropriate blocking buffer which decreases background interference and improves the signal-to-noise ratio. In ELISA once the antibody or protein antigens have been properly adsorbed onto the plate, the next critical step in creating a reliable assay is the blocking of the plate, as this will ensure an accurate signal. In a western blot experiment your membrane has been chosen for the ability to bind protein, thus steps must be taken to prevent interactions between the membrane and the antibody used for detection of the target protein. Blocking of non-specific binding is achieved by placing the membrane in a dilute solution of protein (blocking buffer), with a small percentage of detergent. The protein in the dilute solution attaches to the membrane in all places where the target proteins have not attached. Thus, when the antibody is added, there is no room on the membrane for it to attach other than on the binding sites of the specific target protein. This reduces "noise" in the final product, provides clearer results, and eliminates false positives.
Rockland Immunochemicals Blocking Buffer formulations reduce nonspecific binding of sample and assay components to the well while stabilizing the coated protein. Our solutions can help develop more accurate and long lasting results that drive scientific innovation. Our products are dependable and high quality. Don’t degrade your success. Compromise elsewhere. Use Rockland blocking buffers to boost your results today.
Western Blot or immunoblot is a work horse immunoassay for most labs, used to demonstrate the presence or absence of important proteins, detect post-translational modifications, diagnose disease and more. Robust bands that identify targets can be visualized after probing your western blot with Rockland primary antibodies for detection and conjugated secondary antibodies. Rockland has also developed Revitablot™ Western Blot Stripping Buffer, which contains solutions in a proprietary combination to enhance the removal of bound antibodies from western blot membranes for repeated use. The proprietary formulation of the solution ensures high stripping efficiency with low backgrounds.
Revitablot™ Western Blot Stripping Buffer(500mL) is a uniquely formulated, ready-to-use reagent specifically designed for western blotting. Revitablot™ is faster and more efficient at stripping primary and secondary antibodies, allowing blots to be stripped multiple times for the repeated use of membranes. This solution can be used at room temperature and only requires 5-20 minutes of incubation time to strip the membrane. Revitablot™ Western Blot Stripping Buffer(50mL) is gentle and effective on both nitrocellulose and PVDF western blot membranes.
Rockland phospho blocking buffers reduce the background noise of particular assay types. Rockland remains a leader in blocking buffer development, maintaining continuous quality control and process management throughout production. View our large and diverse set of phospho specific antibodies and reagents that detect phosphorylated targets with great precision.
Common applications for blocking buffers include:
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