Biotinylated Anti-Streptavidin has been widely used as an amplifying reagent in immunohistochemistry, in situ hybridization, microarray assays, ELISAs, blots, and many other applications. The capability of binding streptavidin via either biotin binding sites or through antigen binding sites, makes this biotinylated antibody unique. Anti-Streptavidin does not bind avidin, and Anti-Avidin does not recognize streptavidin.
Immunofluorescence, In situ hybridization
0.5 mg active conjugate/ml
2-8 °C; Store frozen for long term storage
10 mM HEPES, pH 7.5, 0.15 M NaCl, 0.08% sodium azide, 20 mg/ml sucrose.
The recommended concentration range for use is 1-10 µg/ml.If this biotinylated antibody is to be used in tissues, which may contain cross-reacting endogenous
immunoglobulins, dilution of this biotinylated antibody may be made in buffers containing 2% normal serum from the same species as the tissue.
These antibodies can be used either as part of preformed complexes or in sequence to amplify fluorescent signals. When used in sequence, the target is first labeled with fluorochrome-conjugated avidin or streptavidin, followed by incubation with Biotinylated Anti-Avidin or Biotinylated Anti-Streptavidin, followed by a second layer of fluorochrome-conjugated avidin or streptavidin. This sequence can be repeated. This multi-layered approach introduces more fluorochromes at the target site and can provide a multi-fold amplification over a single layer.
These affinity purified antibodies are also available unconjugated or fluorescein-labeled.
The following figures illustrate different experimental setups used to achieve increasing levels of sensitivity.