Immunohistology buffers

Low background and strong specific reaction. Standardized staining.

ISO 9008 certified for diagnostics

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Any antibody preparation has some potential to produce non-specific reaction in the assay. This originates from:

• non-specific antibodies that are present in some proportion in any polyclonal antibody preparation, including affinity purified ones (often "affinity purified" means only isolation of IgG fraction on Prot A/G column, not the purification on the antigen column under very stringent conditions)

• low specificity antibodies among specific ones in polyclonal

• fragments of fallen apart IgGs in stored preparations, including monoclonal

• separate heavy and light chains of specific antibodies, produced by most hybridomas

All these are capable of binding non-specifically to molecules on tissue sections, blots, fixed cells and other objects for immune detection. In case of retrieved formalin sections the risk of non-specific reaction is even higher, since to activate the epitope recovery the proteins comprising the tissue sections are denatured during HIER, thus making many domains accessible that are charged and capable of binding the test immunoglobulins in a non-specific manner.

The standard means to block non-specific binding of specific antibody preparation is to add an irrelevant protein, such as BSA, other serum, casein, etc. However, everyone who tried to do this knows that increasing (for effective blocking) concentration of such blocking agent leads to a great reduction of the specific reaction as well. This is due to large blocking molecules binding to accessible sites on section and thus sterically blocking access of specific antibodies to epitopes of interest (schematically represented in the figure on the left, top). All our buffers developed for immune assays isntead contain short (0.6-2 kD) peptides that are capable of effectively blocking non-specific reactions while not affecting the specific binding of antibody.(fugure on the left, bottom)

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Aptum's collection of Immunohistology buffers has also some other benefits (see below) and allows you to achieve the best quality IHC result without compromising the antigen detection. The buffers can also be used in other immune assays, such as immunofluorescence on sections, flow cytometry on fixed cells, hybridization of sections with antibody detection.

The Retriever IHC buffers empower you to control non-specific staining on every step of immunohistochemistry. They are especially highly recommended for research pathology where, in contrast to diagnostics, many polyclonal and/or low-affinity antibodies are used.