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CA15-3

IHC of CA15-3 on an FFPE Breast Tissue

Description

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.

Antibody Type
Mouse Monoclonal
Clone
DF3
Isotype
IgG1/K
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Breast, Pancreas, Salivary Gland
Storage
Store at 2°-8°C
Stability
2 years

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).

Presentation

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.

Availability
Catalog No.
Antibody Type
Dilution
Volume/QTY
BSB 5099
Prediluted
Ready-To-Use
3.0 ml
BSB 5100
Prediluted
Ready-To-Use
7.0 ml
BSB 5101
Prediluted
Ready-To-Use
15.0 ml
BSB 5102
Concentrated
1:50-1:300
0.1 ml
BSB 5103
Concentrated
1:50-1:300
0.5 ml
BSB 5104
Concentrated
1:50-1:300
1.0 ml
BSB 5105
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Gatalica Z, et al. Applied IHC. 1994;2(3):205-211
  2. Kufe D, et al. Hybridoma. 1984;3:223-32
  3. Colomer R, et al. Breast CA Res Treat. 1989;13:123-33
  4. Ho JJL, et al. Int J Cancer. 1992;52:693-700

Recommended Immunohistochemical Protocol

Pretreatment
  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.

Immunohistochemical Protocol

Step

ImmunoDetector
(AP or HRP)

PolyDetector
(AP or HRP)
Peroxidase/AP Block
5 minutes
5 minutes
Primary Antibody
30 minutes
45 minutes
Secondary Biotinylated Link
10 minutes
Not Applicable
AP or HRP Label
10 minutes
45 minutes
Substrate-Chromogen
5-10 minutes
10 minutes
Counterstaining
Time varies with counterstain
Time varies with counterstain