Compatibility check – Dako Autostainer User Manual

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Compatibility Check


A compatibility check is included to minimize programming errors resulting from
assigning incompatible reagents (such as alkaline phosphatase-labeled streptavidin and
DAB substrate) to a slide protocol. This feature has two check levels. The first check is
for the species reactivity compatibility of the primary antibody and the secondary
reagent. The second addresses the compatibility rules dictated by the enzyme used in
the detection system. The following tables summarize the compatibility rules and codes
assigned to each reagent type.

Every reagent in the reagent categories list (or Protocol Element) can be programmed
with an appropriate compatibility code or combination of codes. For example, a linking
reagent that reacts with mouse and rabbit antibodies can be marked with compatibility
code AB. The software verifies that these compatibility rules are not violated during the
programming of a staining run. If an incompatible reagent is assigned to a tile in the
programming grid, a message displays the two reagents that are incompatible.

Species Reactivity

Reagent Type

(Protocol Element)

Description Compatibility

Code

Primary Antibody

Monoclonal primary antibodies raised in
mouse (e.g., mouse anti-human)

A

Primary Antibody

Polyclonal primary antibodies raised in
rabbit (e.g., rabbit anti-human)

B

Secondary Reagent

Secondary reagents (antibodies)
compatible with monoclonal antibodies
raised in mouse (e.g., biotinylated anti-
mouse IgG)

A

Secondary Reagent

Secondary reagents (antibodies) reacting
with polyclonal antibodies raised in rabbit
(e.g., biotinylated anti-rabbit IgG)

B

Secondary Reagent

Secondary reagents (antibodies) reacting
with both monoclonal and polyclonal
antibodies raised in mouse or rabbit,
respectively

AB


Enzyme System Compatibility

Reagent Type

(Protocol Element)

Description Compatibility

Code

Endogenous Enzyme

Block, HRP-compatible

X

Endogenous Enzyme

Block, AP-compatible

Y

Tertiary Reagent

HRP-labeled streptavidin

X

Tertiary Reagent

AP-labeled streptavidin

Y

Substrate

HRP-compatible substrates (e.g., DAB, AEC) X

Substrate

AP-compatible substrates (e.g., Fast Red,
New Fuchsin)

Y

Autostainer Handbook

29

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