Ventana Molecular Discovery Systems 1849300, Rev. A April 2003 1 http://www.ventanadiscovery.com

Discovery™ Universal Secondary Antibody

Secondary antibody for use with Discovery™ Detection kits on the Ventana Discovery™ Staining Platform

Catalog Number: 760-4205 250 Tests Introduction Discovery Universal Secondary Antibody is designed to work with DAB Map and BlueMap detection kits when mouse or rabbit primary antibodies are employed. The Universal Secondary Antibody can be used for antigen localization in frozen sections or formalin or paraformaldehyde fixed, paraffin-embedded tissue sections.

Kit Components Storage Discovery Universal Secondary Antibody Package Insert 1 copy Discovery Universal Secondary Antibody (For use with mouse or rabbit primary antibodies) 250 tests 2-8°C

Discovery Reagents Required But Not Provided With This Kit

Reagent Catalog # Size Storage DAB Map Detection Kit OR (Horseradish peroxidase DAB detection kit) 760-102 250 tests 2-8°C

BlueMap Detection Kit (Alkaline phosphatase NBT/BCIP detection kit) 760-120 125 tests 2-8°C

Materials Required But Not Supplied By Ventana Medical Systems Formalin-fixed, paraffin-embedded tissue sections, frozen sections or cytological preparations Getting Started Discovery Universal Secondary Antibody The Discovery Universal Secondary Antibody comes ready to use. Register the reagent on your Discovery system and place the dispenser on the reagent wheel. This reagent can be used with any mouse or rabbit primary antibody and therefore should not be used on mouse or rabbit tissue as high “background” will occur due to endogenous immunoglobulin staining. Controls A known positive tissue should be run with every antibody being used. This tissue serves as a reagent control to demonstrate that all of the reagents used in the assay are functional. The staining of the positive control also serves as a baseline for evaluating run to run and/or day to day consistency. In addition to the positive control, a non-immune Ig control should be run on every tissue tested. Thus, the specificity of the staining reaction can be documented by comparing the non-immune Ig staining to the primary antibody staining. Easy Steps for using the Discovery Universal Secondary Antibody within a Protocol

1. To edit an existing protocol to include the Discovery Universal Secondary Antibody, open the NexES software. 2. Click on the “Protocols” button on the main screen. A window will appear on the screen with “Create/Edit Protocol” and “Manage Protocol”.

Click on “Create/Edit Protocol” to open the “NexES Protocol Editor-Discovery Staining Module” window. 3. Select the “Research IHC DAB Map” or appropriate procedure under the “Procedure” field.

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4. Select the appropriate Protocol under the “Protocol” field.

5. Check the box next to the “2nd Antibody” field.

6. Check the box next to “2nd Antibody Autodispense” which now appears on the screen. 7. Two new windows appear on the screen. In the “User Detection” window, select “Universal Secondary Antibody” and in the “Plus Incubation

Time” select 16 minutes.

Saving the Protocol 1. Click on the “Save As” button. A window will appear with fields for a protocol name and protocol number. Type in an unused name for the

protocol and select an unused number in the appropriate boxes. Click on the “Save” button and the protocol will be saved.

Instructions for Use Open the package and take out the dispenser. Remove the cap from the nozzle of the dispenser and place it on the nozzle cap holder on the rear of the dispenser. While holding the dispenser upright, remove the yellow shipping key by pulling the key tab to disengage it from each end. Do not cover the nozzle tip or depress the dispenser while removing key. Place the dispenser on the reagent tray along with the appropriate Ventana Discovery accessory reagents. Bulk reagents should be prepared using quality reagent grade water. (Do not use tap water). Carboys for storing bulk reagents should be rinsed thoroughly between fillings.

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Intended Use

For laboratory research only. Not for in vitro diagnostic use. Technical Consultation Additional technical information can be obtained from our Molecular Discovery Systems Technical Center at:

U.S.A. : (800) 227-2155 Ventana Medical Systems, Inc. 1910 Innovation Park Drive Tucson, AZ 85737 USA

Europe : (+33) 3 90 40 52 00 Ventana Medical Systems S.A . Parc d’innovation, BP 144 Rue G. de Kaysersberg F-67404 ILLKIRCH Cedex France

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Ventana Japan K.K. Landmark Tower, 35F 2-2-1, Minatomirai, Nishi-ku Yokohama, 220-8135 Japan

Australia : +61 (0) 3 9817 3077 Ventana Medical Systems, Pty. Ltd. P.O. Box 743 Deepdene Victoria 3103 Australia