Why blocking solution

Purpose and function of blocking steps Types of blocking buffers Which blocking buffer to use? Due to the number of proteins within milk, milk may mask some antigens and lower the detection limit of the western blot. BSA is generally a weaker blocker, which can result in more non-specific antibody binding, but can increase the detection sensitivity for low-abundant proteins. Purified proteins e. Ideal when blockers, such as non-fat milk, block antigen-antibody binding More expensive than traditional non-fat milk formulations.

Click image to enlarge. Recommended Thermo Scientific western blot blocking buffers. Blocking agent Highlights When to use Available formats Pierce Clear Milk Blocking Buffer Clarified and stabilized milk proteins High-performance replacement for homemade milk blocking buffers Long shelf-life at room temperature Use when high background seen with Non-fat milk Fluorescent and chemiluminescent applications Borate, pH 7.

DenHollander, N, Defus D. J Immunol Methods 1 — Education center homepage. Western blotting protocols. Western blotting webinars. For Research Use Only. Not for use in diagnostic procedures. Sign in. Account Check Order Status. Contains biotin and phosphoproteins, which can interfere with streptavidin-biotin detection strategies and detection of phosphorylated target proteins. Good alternative to milk, can be used in biotin-streptavidin systems or when probing for phosphoproteins.

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. The proprietary formulation of the solution ensures high stripping efficiency with low backgrounds.

It 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.

Rockland phospho blocking buffers reduce the background noise of particular assay types. Martin St. Outlying Islands U. Product Categories. Browse all Blocking Reagents. Explore Blocking Reagents. Protein Blocking Solutions.

Autofluorescence Quencher. Normal serum may have been added as indicated on the label. The blocking capacity of each lot is determined using a dot-spot test system as described by Moeremans et al. Methods 74, , Each package contains 30 ml of solution.