Intended use: Antibody Diluent B is especially developed for dilution of certain primary antibodies. Antibodies diluted with Antibody Diluent B are primarily used in immunohistochemistry with formalin-fixed paraffin-embedded tissue sections, but also with frozen, HOPE-fixed, and cytological samples as well as in immunoblot procedures.
Summary and explanation: Antibody diluents used in immunohistochemistry should protect the antibody from microbial contamination and stabilize the antibody chemically. Antibody Diluent B reduces non-specific binding of antibodies to tissue sections and is therefore extremely useful in receiving background-free staining results.
Reagents provided: 100 ml Antibody Diluent B (ready-to-use)
Storage and handling: The solution should be stored at 2-8°C without furt her dilution. Do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Principle of method: Immunohistochemical staining procedures often start with incubation of a blocking solution to reduce unspecific binding of primary antibody to tissue sections. This step can be omitted if the antibody used is diluted in Antibody Diluent B. Antibody Diluent B • minimises unspecific binding of the primary antibody to the tissue section, • reduces surface tension of the antibody solution and improves spreading the reagent on the slide, • increases microbial and chemical stability of the antibody, • reduces adhesion of antibody to the surface of the vial, • and minimises the danger of antibody degradation by proteolytic enzymes.
Expected results: During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
References: Elias JM Immunohistopathology – A practical Approach to Diagnosis ASCP Press 2003/Nadji M and Morales AR Ann N.Y. Acad Sci 420:134-139, 1983
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