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Product details

SynonymsR3C3, Nuclear receptor subfamily 3 group C member 3, PGR, PR, PRA, PRB, Progesterone receptor form A, Progesterone receptor form B

Antibody type = Recombinant Rabbit monoclonal / IgG

Clone = MSVA-570R

Positive controlUterine cervix: almost all columnar epithelial cells, basal squamous epithelial cells and most of the stromal cells must show a strong nuclear staining with only minimal cytoplasmic staining.

Negative controlTonsil: PR immunostaining should be absent in all epithelial and non-epithelial cells.

Cellular localization = Nucleus

Reactivity = Human

 

Application = Immunohistochemistry
Dilution = 1:50
Intended Use = Research Use Only

Relevance of Antibody

Progesterone Receptor is expressed in progesterone dependent cell types.

Biology Behind

The progesterone receptor (PR), is a nuclear receptor protein coded by the  PGR gene residing on chromosome 11q22. It is activated by the steroid hormone progesterone. In the absence of progesterone its carboxyl terminal inhibits transcription of target genes. After binding to progesterone, a structural change of the PR protein terminates the inhibitory action, the protein dimerizes, enters the nucleus, binds to DNA and induces transcription of target genes. Progesterone antagonists prevent the structural reconfiguration. Progesterone and its receptor are key elements of female reproduction. In breast development, progesterone is involved in the formation of lobular-alveolar units. In human endometrium, progesterone directs glandular differentiation, stromal proliferation and development of decidual cells. PR shares considerable sequence homology with the estrogen receptor (ER) in the DNA-binding central domain. The presence of a functional ER is required for PR synthesis in the cell. PR exists in two isoforms, PR-A and PR-B, which are often co-expressed in human tissues. They are transcribed from two different promoters of the PGR gene. The two PR isoforms are identical except that PR-A lacks 164 amino acids contained at the N-terminal end of PR-B. To some extent, the two isoforms have antagonistic functions. Predominant expression of one isoform can occur in some tissues under specific circumstances. PR-A is the major isoform in the uterine stroma, and PR-B is the predominant isoform in the endometrial glands. Normal human breast expresses PR-A and PR-B to equal extent.

Staining Pattern in Normal Tissues

Progesterone Receptor staining pattern in Normal Tissues with antibody MSVA-570R (images are shown in our “Normal Tissue Gallery”)

Brain Cerebrum Negative.
Cerebellum Negative.
Endocrine Tissues Thyroid Negative.
Parathyroid Negative.
Adrenal gland Weak to moderate PR staining of a variable fraction of adrenocortical cells (not in all samples).
Pituitary gland Weak to moderate PR staining of a variable number of epithelial cells in the adenohypophysis. Staining is absent in the neurohypophysis.
Respiratory system Respiratory epithelium Negative.
Lung Negative.
Gastrointestinal Tract Salivary glands Weak to moderate PR positivity of some cells.
Esophagus Weak to moderate PR positivity of some muscular cells in the muscular wall.
Stomach Negative.
Duodenum Weak to moderate PR positivity of some cells in Brunner glands.
Small intestine Weak to moderate PR positivity of some muscular cells in the muscular wall.
Appendix Negative.
Colon Negative.
Rectum Negative.
Liver Negative.
Gallbladder Negative.
Pancreas Weak to moderate to strong PR staining of a large fraction of islet cells.
Genitourinary Kidney Weak to moderate PR staining in subsets of glomerular, tubular and stromal cells.
Urothelium Weak to moderate to strong PR staining in stroma cells of the urinary bladder and the pyelon. The urothelium is negative.
Male genital Prostate Moderate to strong PR staining of stromal cells.
Seminal vesicles Moderate to strong PR staining of stromal cells.
Testis Negative.
Epididymis Weak to moderate PR staining of epithelial cells of the cauda.
Female genital Breast Moderate to strong PR staining of a fraction of luminal epithelial cells.
Uterus, myometrium Strong PR staining of muscular cells.
Uterus, ectocervix Strong PR staining of stromal cells and basal squamous epithelial cells.
Uterus endocervix Strong PR staining of epithelial and stromal cells.
Uterus, endometrium Strong PR staining of epithelial, decidual, and stromal cells.
Fallopian Tube Strong PR staining of epithelial and stromal cells.
Ovary Strong PR staining of stromal cells and corpus luteum.
Placenta early Negative.
Placenta mature Negative.
Amnion Negative.
Chorion Negative.
Skin Epidermis Negative.
Sebaceous glands Negative.
Muscle/connective tissue Heart muscle Negative.
Skeletal muscle Negative.
Smooth muscle Negative.
Vessel walls Moderate to strong PR staining of smooth muscle cells in the media of the aorta.
Fat Negative.
Stroma PR staining can be found in some stroma cells of various organs.
Endothelium Negative.
Bone marrow/ lymphoid tissue Bone marrow Negative.
Lymph node Negative.
Spleen Negative.
Thymus Negative.
Tonsil Negative.
Remarks PR staining is nuclear.

 

These findings are largely consistent with the RNA and protein data described in the Human Protein Atlas (Tissue expression Progesterone Receptor) 

 

Positive control: Uterine cervix: almost all columnar epithelial cells, basal squamous epithelial cells and most of the stromal cells must show a strong nuclear staining with only minimal cytoplasmic staining.

Negative control: Tonsil: PR immunostaining should be absent in all epithelial and non-epithelial cells.

Normal tissue gallery

Staining Pattern in Relevant Tumor Types

The TCGA database on RNA expression in cancer has described the highest levels of PR expression in breast and endometrial cancer followed by ovarian and cervical cancer. Most other important tumor entities are described to be usually “PR negative”.

The TCGA findings on Progesterone Receptor RNA expression in different tumor categories have been summarized in the Human Protein Atlas.

 

 

 

Cancer tissue gallery

Compatibility of Antibodies

No data available at the moment

Protocol Recommendations

IHC users have different preferences on how the stains should look like. Some prefer high staining intensity of the target stain and even accept some background. Others favor absolute specificity and lighter target stains. Factors that invariably lead to more intense staining include higher concentration of the antibody and visualization tools, longer incubation time, higher temperature during incubation, higher temperature and longer duration of the heat induced epitope retrieval (slide pretreatment). The impact of the pH during slide pretreatment has variable effects and depends on the antibody and the target protein.

 

All images and data shown here and in our image galleries are obtained by the manual protocol described below. Other protocols resulting in equivalent staining are described as well.

 

Manual protocol

Freshly cut sections should be used (less than 10 days between cutting and staining). Heat-induced antigen retrieval for 5 minutes in an autoclave at 121°C in pH 7,8 Target Retrieval Solution buffer. Apply MSVA-570R at a dilution of 1:50 at 37°C for 60 minutes. Visualization of bound antibody by the EnVision Kit (Dako, Agilent) according to the manufacturer’s directions.

 

 

 

Agilent / Dako – Autostainer Link 48

Pretreatment in PT-Link for 30 minutes at 95°C (pH high); FLEX peroxidase blocking for 5 minutes (room temperature), MSVA-570R 1:50 for 20 minutes (room temperature), FLEX+ mouse/rabbit (LINKER) for 15 minutes (room temperature), horseradish peroxidase (HRP) for 20 minutes (room temperature), FLEX DAB+Sub-Chromo for 10 minutes (room temperature), FLEX hematoxylin for 5 minutes (room temperature).

These images reflect stainings by the protocol described above. It is of note that a comparable staining result can also be obtained by different protocols. In general, a longer pretreatment, a longer incubation time of the primary antibody, a higher antibody concentration, and a longer incubation time of FLEX+LINKER result in stronger staining, potentially at the cost of more background staining. Modifications of the protocol with a strengthening effect on staining intensity in combination with changes of other parameters that result in lower staining intensity can result in a comparable result as shown above.

 


Leica – BOND RX

Dewax at 72°C for 30 seconds; Pretreatment in Bond Epitope Retrieval Solution (ER2 – EDTA pH9) for 20 minutes at 100°C; Peroxidase blocking for 5 minutes (room temperature), MSVA-570R 1:100 for 15 minutes (room temperature), Post primary (rabbit anti mouse) for 8 minutes (room temperature), Polymer (goat anti rabbit) for 8 minutes (room temperature), mixed DAB refine for 10 minutes (room temperature), hematoxylin for 5 minutes (room temperature).

These images reflect stainings by the protocol described above. It is of note that a comparable staining result can also be obtained by different protocols. In general, a longer pretreatment, a longer incubation time of the primary antibody, a higher antibody concentration, a higher temperature during incubation, and a longer incubation time of Post primary and or the Polymer result in stronger staining, potentially at the cost of more background staining. Modifications of the protocol with a strengthening effect on staining intensity in combination with changes of other parameters that result in lower staining intensity can result in a comparable result as shown above.

 

 

Roche – Ventana Discovery ULTRA

Pretreatment for 64 minutes at 100°C (pH 8,4); CM peroxidase blocking for 12 minutes (room temperature), MSVA-570R 1:50 for 20 minutes at 36°C, secondary antibody (anti-rabbit HQ) for 12 minutes at 36°C, anti-HQ HRP for 12 minutes at room temperature, DAB at room temperature, hematoxylin II at room temperature for 8 minutes, bluing reagent at room temperature for 4 minutes.

 

These images depict staining results obtained by the protocol described above. It is of note, that the Ventana machines generally require higher antibody concentrations than other commonly used autostainers because the antibodies are automatically diluted during the procedure. Various other protocols can result in an identical result as shown above. A longer pretreatment, a longer incubation time of the primary antibody, a higher antibody concentration, a higher temperature during incubation, and a longer incubation time of secondary antibody and or the anti-HQ HRP result in stronger staining, potentially at the cost of more background staining.

Potential Research Applications

  • The diagnostic utility of PR expression analysis should be investigated in a large cohort of tumors from different entities.
  • The clinical/biological significance of the rare ER-/PR+ breast cancers is unclear.
  • Much is still unknown on the site-specific role of PR in various different tissues.