
This antibody has been used for evaluating the primary site of a metastatic carcinoma of unknown origin and distinguishing between benign and malignant lesions. It is believed that CA15-3 reacts primarily with the DF3-antigen, a 300 kDa mucin-like glycoprotein present on the apical border of secretory mammary epithelial cells.
CA15-3 has been detected with immunohistochemistry in a wide spectrum of carcinomas, including Breast Carcinomas (ductal and lobular), Sarcomas (Synovial Sarcoma and Malignant Fibrous Histiocytomas), and Lung Carcinomas. CA15-3 can be used as a supplementary marker for epithelial differentiation. CA15-3 does not stain Melanomas or Ewing’s Sarcomas. Approximately 30% of Hepatocellular Carcinomas are positive for CA15-3.
For long-term storage of the concentrated antibody, it is recommended that aliquots of the antibody be frozen at -20°C in glycerol 50% (frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Dilute using an antibody diluent such as ImmunoDetector Protein Block/Antibody Diluent (BSB 0040 and BSB 0041) or ImmunoDNA Background Blocker (BSB 0103-BSB 0107).
CA15-3 is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.
- Gatalica Z, et al. Applied IHC. 1994;2(3):205-211
- Kufe D, et al. Hybridoma. 1984;3:223-32
- Colomer R, et al. Breast CA Res Treat. 1989;13:123-33
- Ho JJL, et al. Int J Cancer. 1992;52:693-700
Recommended Immunohistochemical Protocol
- Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
- Air dry for 2 hours at 58° C.
- Deparaffinize, dehydrate and rehydrate tissues.
- Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
- Any of three heating methods may be used:
- Electric Pressure Cooker
Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature. - Water Bath Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes. - 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.
- Electric Pressure Cooker
- After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
- Wash slides with IHC wash buffer or DI water.
- Continue IHC staining protocol.
Immunohistochemical Protocol
ImmunoDetector
(AP or HRP)
(AP or HRP)