BCA-225

This antibody recognizes a human Breast Carcinoma-associated glycoprotein, BCA-225 (220-225 kDa). This protein differs in size and distribution from other Breast Carcinoma antigens. Unlike other carcinoma antibodies against Breast Carcinoma antigens, this antibody does not react with benign or malignant colonic tissues. Since this antigen is localized in malignancies of Breast Carcinomas and Carcinoma of the Uterine Cervix, it can be effectively used to identify metastatic Breast Carcinoma lesions.

Strong intracytoplasmic staining is seen in primary and metastatic Breast Carcinoma tissue, as well as in Cervical Carcinomas. Apical staining is seen in normal kidney, lung, Fallopian tube, liver, skin (eccrine sweat glands) and uterus. Similar staining patterns are observed in lung, ovarian, and endometrial cancers. Carcinomas of the colon, stomach, prostate, urinary bladder, liver, pancreas, thyroid, and parotid are negative, as are Sarcomas and Lymphoid Cancers.

BCA-225 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.

1. Ceriani RL, Monoclonal Ab’s and Breast Cancer. Boston, Martinus, Nijhoff. 1985
2. Mesa-Tejada R, et al. Am J Pathol. 1988;130:305-314

1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
2. Air dry for 2 hours at 58° C.
3. Deparaffinize, dehydrate and rehydrate tissues.
4. 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).
5. 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.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
7. Wash slides with IHC wash buffer or DI water.
8. Continue IHC staining protocol.