CA15-3 Antibody – Mouse Monoclonal

This antibody is intended for use in Immunohistochemical applications on formalin-fixed paraffin-embedded tissues (FFPE), frozen tissue sections and cell preparations.

Specimen Preparation:
Paraffin sections: The antibody can be used on formalin-fixed paraffin-embedded (FFPE) tissue sections. Ensure tissue undergoes appropriate fixation for best results. Pre-treatment of tissues with heat-induced epitope retrieval (HIER) is recommended using ImmunoDNA Retriever with Citrate , ImmunoDNA Retriever with EDTA or ImmunoDNA Digestor. See reverse side for complete protocol. Tissue should remain hydrated via use of Immuno/DNA Washer solutions.
Frozen sections and cell preparations: The antibody can be used for labeling acetone-fixed frozen sections and acetone-fixed cell preparations.

Staining Procedure:
1. Cut and mount 3-5 micron formalin-fixed paraffin-embedded tissues on positively charged slides such as Hydrophilic Plus Slides.
2. Air dry for 2 hours at 58°C.
3. Deparaffinize, dehydrate and rehydrate tissues.
4. Subject tissues to heat induced epitope retrieval (HIER) using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate or EDTA.
5. Any of three heating methods may be used:
a. TintoRetriever Pressure Cooker or Equivalent
Place tissues/slides in a staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA, and place on trivet in the pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high. Incubate for 15 minutes. Open and immediately transfer slides to room temperature.
b. TintoRetriever PT Module or Water Bath Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA at 95°-99°C. Incubate for 30-60 minutes.
c. Conventional Steamer Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a steamer, cover and steam for 30-60 minutes.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
7. For manual staining, perform antibody incubation at ambient temperature. For automated staining methods, perform antibody incubation according to instrument manufacturer’s instructions.
8. Wash slides with ImmunoDNA washer or DI water.
9. Continue IHC staining protocol. Wash slides between each step with ImmunoDNA washer solution.

Product Limitations: Due to inherent variability present in immunohistochemical procedures (including fixation time of tissues, dilution factor of antibody, retrieval method utilized and incubation time), optimal performance should be established through the use of positive and negative controls.