Recombinant Anti-Progesterone Receptor antibody [YR85] (ab32085)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [YR85] to Progesterone Receptor
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Progesterone Receptor antibody [YR85]
See all Progesterone Receptor primary antibodies -
Description
Rabbit monoclonal [YR85] to Progesterone Receptor -
Host species
Rabbit -
Specificity
This antibody recognises progesterone receptor. The antibody does not cross-react with other NR3 family members. Since the recognized epitope is near the N-terminal end of the protein, this product should only detect isoform B and not isoform A.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Progesterone Receptor (N terminal). The exact sequence is proprietary.
Database link: P06401 -
Positive control
- Human breast carcinoma, T47D cell lysate. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: DU145.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Dissociation constant (KD)
KD = 2.48 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
YR85 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32085 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
1/1000 - 1/10000. Predicted molecular weight: 99 kDa.
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IHC-P | (4) |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP |
1/150.
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ICC/IF |
Use a concentration of 0.2 - 5 µg/ml.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/1000 - 1/10000. Predicted molecular weight: 99 kDa. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
1/150. |
ICC/IF
Use a concentration of 0.2 - 5 µg/ml. |
Target
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Function
The steroid hormones and their receptors are involved in the regulation of eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Progesterone receptor isoform B (PRB) is involved activation of c-SRC/MAPK signaling on hormone stimulation.
Isoform A: inactive in stimulating c-Src/MAPK signaling on hormone stimulation.
Isoform 4: Increases mitochondrial membrane potential and cellular respiration upon stimulation by progesterone. -
Sequence similarities
Belongs to the nuclear hormone receptor family. NR3 subfamily.
Contains 1 nuclear receptor DNA-binding domain. -
Domain
Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain. -
Post-translational
modificationsPhosphorylated on multiple serine sites. Several of these sites are hormone-dependent. Phosphorylation on Ser-294 occurs preferentially on isoform B, is highly hormone-dependent and modulates ubiquitination and sumoylation on Lys-388. Phosphorylation on Ser-102 and Ser-345 also requires induction by hormone. Basal phosphorylation on Ser-81, Ser-162, Ser-190 and Ser-400 is increased in response to progesterone and can be phosphorylated in vitro by the CDK2-A1 complex. Increased levels of phosphorylation on Ser-400 also in the presence of EGF, heregulin, IGF, PMA and FBS. Phosphorylation at this site by CDK2 is ligand-independent, and increases nuclear translocation and transcriptional activity. Phosphorylation at Ser-162 and Ser-294, but not at Ser-190, is impaired during the G(2)/M phase of the cell cycle. Phosphorylation on Ser-345 by ERK1/2 MAPK is required for interaction with SP1.
Sumoylation is hormone-dependent and represses transcriptional activity. Sumoylation on all three sites is enhanced by PIAS3. Desumoylated by SENP1. Sumoylation on Lys-388, the main site of sumoylation, is repressed by ubiquitination on the same site, and modulated by phosphorylation at Ser-294.
Ubiquitination is hormone-dependent and represses sumoylation on the same site. Promoted by MAPK-mediated phosphorylation on Ser-294.
Palmitoylated by ZDHHC7 and ZDHHC21. Palmitoylation is required for plasma membrane targeting and for rapid intracellular signaling via ERK and AKT kinases and cAMP generation. -
Cellular localization
Nucleus. Cytoplasm. Nucleoplasmic shuttling is both homone- and cell cycle-dependent. On hormone stimulation, retained in the cytoplasm in the G(1) and G(2)/M phases; Mitochondrion outer membrane and Nucleus. Cytoplasm. Mainly nuclear. - Information by UniProt
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Database links
- Entrez Gene: 5241 Human
- Omim: 607311 Human
- SwissProt: P06401 Human
- Unigene: 32405 Human
- Unigene: 742403 Human
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Alternative names
- NR3C3 antibody
- Nuclear receptor subfamily 3 group C member 3 antibody
- PGR antibody
see all
Images
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Immunofluorescence staining of Recombinant Anti-Progesterone Receptor antibody [YR85] (ab32085) in T47D cells (+ve control) and HepG2 cells (-ve control). The cells were fixed with 100% Methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab32085 at 0.2 µg/mL dilution and ab7291, Anti-alpha Tubulin antibody [DM1A], at 1/1000 dilution, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150087) at 1/1000 (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with the Perkin Elmer Operetta and a maximum intensity projection of confocal planes is shown.
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Immunofluorescence staining of Recombinant Anti-Progesterone Receptor antibody [YR85] (ab32085) in T47D cells (+ve control) and HepG2 cells (-ve control). The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab32085 at 0.2 µg/mL dilution and ab7291, Anti-alpha Tubulin antibody [DM1A], at 1/1000 dilution, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150087) at 1/1000 (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with the Perkin Elmer Operetta and a maximum intensity projection of confocal planes is shown.
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Immunohistochemical analysis of progesterone receptor expression in paraffin embedded human breast carcinoma, using 1/100 ab32085.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Intracellular Flow Cytometry analysis of T47D (human mammary gland ductal carcinoma) cells labeling Progesterone Receptor with purified ab32085 at 1/1100 dilution (1ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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Anti-Progesterone Receptor antibody [YR85] (ab32085) at 1/1000 dilution + Human breast cancer lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution
Predicted band size: 99 kDa
Observed band size: 118 kDa why is the actual band size different from the predicted?
Exposure time: 180 seconds -
Anti-Progesterone Receptor antibody [YR85] (ab32085) at 1/2000 dilution + T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 99 kDa
Observed band size: 118 kDa why is the actual band size different from the predicted?
Exposure time: 40 secondsBlocking buffer: 5% NFDM/TBST.
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ICC/IF image of ab32085 stained DU145 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab32085 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (29)
ab32085 has been referenced in 29 publications.
- Zhang L et al. Progesterone receptor distribution in the human hypothalamus and its association with suicide. Acta Neuropathol Commun 12:16 (2024). PubMed: 38263257
- De Vargas Roditi L et al. Single-cell proteomics defines the cellular heterogeneity of localized prostate cancer. Cell Rep Med 3:100604 (2022). PubMed: 35492239
- Lin YK et al. SCM-198 Prevents Endometriosis by Reversing Low Autophagy of Endometrial Stromal Cell via Balancing ERα and PR Signals. Front Endocrinol (Lausanne) 13:858176 (2022). PubMed: 35784569
- Zhang YL et al. Progesterone suppresses the progression of colonic carcinoma by increasing the activity of the GADD45a/JNK/c-Jun signalling pathway. Oncol Rep 45:N/A (2021). PubMed: 33846816
- Ma X et al. Fatostatin reverses progesterone resistance by inhibiting the SREBP1-NF-?B pathway in endometrial carcinoma. Cell Death Dis 12:544 (2021). PubMed: 34039951