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p63

IHC of p63 on an FFPE Prostate Tissue

Description

In addition to p53, mammalian cells contain two homologous genes, p63 and p73. These genes give rise to the expression of proteins that are highly similar to p53 in structure and function. In particular, p63 and p73 proteins can induce p53-responsive genes and elicit programmed cell death. p73 and p63 are more important during development and differentiation. In particular, p63 appears to be primarily implicated in epithelial development.


Anti-p63 to human p63 protein labels an epitope common to all six p63 isotypes (TAp63α, TAp63β, TAp63γ, ΔNp63α, ΔNp63β, ΔNp63γ). p63 labels the nuclei of myoepithelial cells in the prostate gland as well as breast tissue, making it useful in differentiating benign vs. malignant prostate lesions and breast lesions.

Antibody Type
Mouse Monoclonal
Clone
4A4
Isotype
IgG2a/K
Reactivity
Paraffin, Frozen
Localization
Nuclear
Control
Normal Prostate, Breast
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

p63 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 5848
Prediluted
Ready-To-Use
3.0 ml
BSB 5849
Prediluted
Ready-To-Use
7.0 ml
BSB 5850
Prediluted
Ready-To-Use
15.0 ml
BSB 5851
Concentrated
1:100-1:500
0.1 ml
BSB 5852
Concentrated
1:100-1:500
0.5 ml
BSB 5853
Concentrated
1:100-1:500
1.0 ml
BSB 5854-1
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Yang A, et al. Mol Cell. 1998;2:305-16
  2. Signoretti S, et al. Am J Pathol. 2000;157:1769-75
  3. Yang A, et al. Nature. 1999;398:714-18
  4. Barbareschi M, et al. Am J Surg Pathol. 2001;Aug;25(8);1054-60
  5. Werling RW, et al. Am J Surg Pathol. 2003;Jan;27(1):82-90
  6. Rajal B Shah, et al. Am J Surg Pathol. 2002;26(9):1161-1168
  7. Di Como CJ, et al. Clinical Cancer Research. 2002;Vol.8:494-501
  8. Weinstein MH, et al. Mod Pathol. 2002;Dec;15(12):1302-8

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