Progesterone Receptor (PR) Antibody Staining Protocol for Immunohistochemistry


Description: The role of steroid hormone receptors in breast cancer is well known. The absence of ER and PR predicts early recurrence and poor survival of breast cancer patients. Also, the presence of ER and PR in tumors predicts the potential for benefit from tamoxifen and other endocrine-related therapies. Measurement of ER and PR can be determined semi-quantitatively using IHC or quantitatively using DCC or EIA. Correlation between the semi-quantitative and quantitative evaluations of PR have ranged from 73 to 91% depending on the laboratory and antibody used.


Primary Antibody

Name: Progesterone Receptor (PR) Antibody

Clone: PgR 636, Mouse anti-Human

Supplier: Dako

Catalog Number: M3569

Dilution: 1:400 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Nuclear
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections (no antigen retrieval needed)
Positive Control: Uterine gland cells, breast, prostate, endometrium
Negative Control: Omit primary antibody, isotype control, absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary



1. Charafe-Jauffret E, et al (2004) Immunophenotypic analysis of inflammatory breast cancers: identification of an inflammatory signature. J Pathology. 202:265-273.