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Lab Standard Operating Protocol (SOP) for Tyramide Amplification
of Streptavidin-Horseradish peroxidase immunohistochemistry
Materials
Literature
Follow routine Streptavidin-HRP IHC ptotocol for cells or tissue
up to streptavidin step.
11a. Incubate slides for 3-10 minutes in working Biotinyl Tyramide
Amplification Reagent. (Perkin Elmer Life Science Products, product
# NEL700A)
To make working solution of Amplification Reagent dilute the Biotinyl
Tyramide Stock Solution 1:50 using 1X Amplification Diluent. Use
approx. 100-300 ul of working solutin per slide and discard any
unused portion in an appropriate manner. I begin with 5 min. and
increase or decrease the time in subsequent procedures after seeing
the stained slide (the reagent uses HRP to catalyze deposition
of biotin labeled tyramide to enhance signal).
11b.
Wash slides 3X in buffer.
12. Apply label (Streptavadin, Dako , LSAB+ Kit K0690) for 30 min.
at RT
Proceed
as per other IHC protocols
Last
GLP modification 8/26/04:
Jennifer Casati
James R. Turk
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