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Progesterone Receptor

IHC of Progesterone Receptor on an FFPE Breast Carcinoma Tissue

Description

The progesterone receptor (PR) also known as NR3C3 (nuclear receptor subfamily 3, group C, member 3), is an intracellular steroid receptor that specifically binds progesterone. PR is encoded by a single gene PGR residing on chromosome 11q22; it has two main forms, A and B, which differ in their molecular weight. Like all steroid receptors, the progesterone receptor has an amino and a carboxyl terminal, and between them the regulatory domain, a DNA binding domain, the hinge section, and the hormone binding domain.


It has been proposed that expression of PR determination indicates a responsive estrogen receptor (ER) pathway, and therefore, may predict likely response to endocrine therapy in human breast cancer. A number of studies have shown that PR determination provides supplementary information to ER in predicting response to endocrine therapy as well as estimating survival. PR has proved superior to ER as a prognostic indicator in some studies.

Antibody Type
Rabbit Monoclonal
Clone
RBT22
Isotype
IgG
Reactivity
Paraffin, Frozen
Localization
Nuclear
Control
Breast Carcinoma
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

Progesterone receptor clone RBT22 is a rabbit 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 5882
Prediluted
Ready-To-Use
3.0 ml
BSB 5883
Prediluted
Ready-To-Use
7.0 ml
BSB 5884
Prediluted
Ready-To-Use
15.0 ml
BSB 5885
Concentrated
1:50-1:250
0.1 ml
BSB 5886
Concentrated
1:50-1:250
0.5 ml
BSB 5887
Concentrated
1:50-1:250
1.0 ml
BSB 5888
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Dabbs DJ, et al. Diagnostic Immunohistochemistry. 2002
  2. Kell DL, et al. Applied Immunohistochemistry. 1993;1(4):275-81
  3. Leong ASY, et al. Applied Immunohistochemistry. 1993;1(4):282-288
  4. Tesch M, et al. Am J Clin Pathol. 1993;99:8-12
  5. Clarke CL, et al. Endocrinology. 1987;121:1123-32
  6. Feil PD, et al. Endocrinology. 1998;123: 2506-13

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