Defining primary antibodies, Assigning positive and negative control reagents – Dako Autostainer User Manual

Page 28

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4. Select Yes and, if the program is new, a dialog box prompts “Save program on

disk?” Otherwise, the Slide Layout Map screen appears. Select Back to Grid to
return to the Programming Grid and the missing rinse appears in the grid.

Defining Primary Antibodies

Each primary antibody reagent is programmed individually and will appear in the
Programming Grid with its programmed pretreatment (see Reagent Management,
Section 7). Primary antibodies and/or pretreatments can be manual steps and are noted
on the Programming Grid and printouts. If a reagent is a manual step (i.e., overnight
incubation, target retrieval, etc.), then it is performed before the slides are loaded.

Note: A primary antibody’s pretreatment automatically appears with the primary antibody
only if the protocol template contains a pretreatment protocol element (see Designing a
Protocol, Section 5).

1. Enter slide information, select a protocol template, and add necessary detection

reagents. When the first programmable Primary Antibody is highlighted, a list of
available Primary Antibodies for the step is displayed.

2. Select the desired antibody from the list using the UP and DOWN arrow keys on the

keyboard and press ENTER.

Note: To make a pretreatment a manual step, right-click on a pretreatment. The list
of available pretreatments is displayed. Select the desired pretreatment from the list.

3. Universal negative controls and previously assigned and saved matched negative

controls are listed at the end of the primary antibody list and are accessed on the
keyboard by pressing the SHIFT key and typing “_”. Once the correct negative
control is selected, press ENTER and the selected negative reagent and
pretreatment will appear in the Programming Grid.

4. Select the Next button, the system checks the grid for missing rinses. If a rinse is

missing between two reagents, a message asks if you want to continue.

5. Select Yes and, if the program is new, a dialog box prompts “Save program on

disk?” Otherwise, the Slide Layout Map screen appears. Select No to return to the
Programming Grid and the missing rinse appears on the screen.

Assigning Positive and Negative Control Reagents

Positive and negative control reagents can be automatically added for each primary
antibody that is added to the Programming Grid. The controls are added as new slides
and contain the matching detection reagents of the test specimen.






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Autostainer Handbook

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