Blocking buffer

Blocking buffers are utilized in immunoassays and other applications such as immunoblotting or immunohistochemistry in order to minimise signal noise generated by non-specific binding in the assay, sensitivity is improved by utilizing an appropriate blocking buffer which decreases background interference and improves the signal-to-noise ratio.

When developing ELISA assays, blocking is a critical step in creating a reliable procedure as this will ensure an accurate signal is received. In an immunoblotting procedure the membrane has to be exposed to blocking buffer 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 blocking buffer with a small percentage of detergent. This ensures that when the antibody of interest 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 leads to a significant reduction in signal noise as well as increasing the chances of eliminating false positives from your results.

Fitzgerald Industries offers a range of detergents and buffers such as PBS for use in developing your own blocking buffers as well as a Universal IHC Diluent/Blocker/Stabilizer for use in Immunohistochemical applications, visit our homepage today to see our full range.