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Beta-Catenin

IHC of Beta-Catenin on an FFPE Breast Tissue

Description

Beta-Catenin is a subunit of the Cadherin protein complex. Cadherins are a type of protein normally expressed on the surface of certain cells. Specifically, Beta Cateinin is a 92 kDa protein normally found in the cytoplasm of the cell in the sub-membranous location. This protein is associated with E-Cadherin and may be essential for the function of E-Cadherin.


Mutations in the Beta-Catenin gene result in the nuclear accumulation of this protein. Nuclear accumulation of this protein has been demonstrated in Fibromatosis lesions of the breast and abdomen, and therefore is useful in differentiating this lesion from other spindle-cell lesions that may occur in these locations.

Antibody Type
Mouse Monoclonal
Clone
14
Isotype
IgG1
Reactivity
Paraffin, Frozen
Localization
Nuclear
Control
Breast, Abdomen
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

Beta-Catenin 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 5085
Prediluted
Ready-To-Use
3.0 ml
BSB 5086
Prediluted
Ready-To-Use
7.0 ml
BSB 5087
Prediluted
Ready-To-Use
15.0 ml
BSB 5088
Concentrated
1:50-1:200
0.1 ml
BSB 5089
Concentrated
1:50-1:200
0.5 ml
BSB 5090
Concentrated
1:50-1:200
1.0 ml
BSB 5091
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Alman BA, et al. Am J Pathol. 1997;Aug.151(2):329-34
  2. Li C, et al. Am J Pathol. 1998;Sep.153(3):709-14
  3. Kuhnen C, et al. Paqthol Rex Pract. 2000;196(5):299-304
  4. Bracke ME, Van Roy FM, Mareel MM, 1996;213(Pt1):123

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