Overview

  • Product name
    Anti-Progesterone Receptor antibody [YR85]
    See all Progesterone Receptor primary antibodies
  • Description
    Rabbit monoclonal [YR85] to Progesterone Receptor
  • Host species
    Rabbit
  • Specificity
    ab32085 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.
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Progesterone Receptor (N terminal). The exact sequence is proprietary.

  • Positive control
    • Human breast carcinoma, T47D cell lysate. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: DU145.
  • General notes

     

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
WB 1/1000 - 1/10000. Predicted molecular weight: 99 kDa.
IHC-P 1/100 - 1/250.
Flow Cyt Use at an assay dependent concentration.
IP 1/150.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • 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
    modifications
    Phosphorylated 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
  • Database links
  • Alternative names
    • NR3C3 antibody
    • Nuclear receptor subfamily 3 group C member 3 antibody
    • PGR antibody
    • PR antibody
    • PRA antibody
    • PRB antibody
    • PRGR_HUMAN antibody
    • Progesterone receptor antibody
    • Progestin receptor form A antibody
    • Progestin receptor form B antibody
    see all

Images

  • Immunohistochemical analysis of progesterone receptor expression in paraffin embedded human breast carcinoma, using 1/100 ab32085.
  • 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 Fluor® 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.

  • 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.

  • Anti-Progesterone Receptor antibody [YR85] (ab32085) at 1/10000 dilution + T47D cell lysate

    Predicted band size: 99 kDa
    Observed band size: 135,99 kDa
    why is the actual band size different from the predicted?

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Guo C  et al. microRNA-10b expression and its correlation with molecular subtypes of early invasive ductal carcinoma. Exp Ther Med 15:2851-2859 (2018). Read more (PubMed: 29599829) »
  • Xie H & Wang H PRL-3 promotes breast cancer progression by downregulating p14ARF-mediated p53 expression. Oncol Lett 15:2795-2800 (2018). Read more (PubMed: 29435006) »
See all 17 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (Uterus)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate ph6.0
Permeabilization
No
Specification
Uterus
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 2% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 27 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Mammary epithelial cells)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citrate buffer
Permeabilization
No
Specification
Mammary epithelial cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Formaldehyde

Mrs. Katie Elliott

Verified customer

Submitted Jul 15 2016

Answer

Es tut mir leid, dass der Antikörper nicht so funktioniert hat wie auf dem Datenblatt angegeben. Ich habe unsere Finanzabteilung beauftragt, Gutschriften für Sie auszustellen. Sie können die Gutschriften auf folgende Weise verwenden:

(1) Gegen die Rechnung dieses Antikörpers verwenden falls diese noch nicht bezahlt wurde
(2) Aufbewahren und gegen eine zukünftige Rechnung verwenden
(3) Wir stellen Ihnen gerne eine komplette Rückerstattung aus, wenn keine offenen Rechnungen mehr ausstehen

Um eine Rückerstattung des Kaufpreises zu erhalten oder um zu bestätigen wie Sie diese Gutschrift benutzen möchten, möchte ich Sie bitten, sich mit unserer Finanzabteilung in Verbindung zu setzen.
Sie können unsere Finanzabteilung per Email unter der Adresse creditcontrol@abcam.com oder telefonisch unter der Nummer 030 896 779 154 erreichen.

Bitte geben Sie die Referenznummer Ihrer Gutschrift immer mit an.

Die Referenznummer ist für Ihre Unterlagen bestimmt. Wir schicken Ihnen die eigentliche Gutschrift per Post zu. Sie enthält auch die Gutschriftnummer, die mit dem Buchstaben CGB beginnt.

Ich hoffe dass Sie trotz dieser schlechten Erfahrung weiterhin Kunde bei uns bleiben. Bitte zögern Sie nicht sich wieder an uns zu wenden, falls Sie weitere Fragen haben.

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Answer

Wir testen unsere Chargen immer, bevor wir unser Datenblatt aktualisieren, und uns ist bei diesem Antikörper aufgefallen, dass die Spezies- Reaktivität in Maus und Ratte eben nicht konstant, bzw gar nicht mehr gegeben war, so dass wir Maus und Ratte als "reagiert nicht" deklariert haben. Seitdem testen wir die beiden Spezies übrigens auch gar nicht mehr.
Warum dies selbst bei einem monoklonalem (!) Antikörper passieren kann, ist uns leider auch nicht bekannt.

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Answer

Vielen Dank für Ihre Email.

Wir haben das Datenblatt im März 2011 geändert, allerdings habe ich keine Hinweise darauf gefunden, dass wir eine Kampagne gestartet haben. Ich habe dies jetzt nachgeholt.

Wenn Sie mir bitte mitteilen, wann Sie denn Antikörper gekauft haben (ungefähres Datum reicht), kann ich ihnen schon jetzt entweder eine Gutschrift, eine Erstattung oder einen kostenlosen Erstsatz zukommen lassen.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (human breast cancer xenograft (MCF7))
Specification
human breast cancer xenograft (MCF7)
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH6.0
Permeabilization
No
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 26 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (breast cancer)
Specification
breast cancer
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate - 20 minutes 100oC
Permeabilization
No

Abcam user community

Verified customer

Submitted Feb 09 2009

Application
ChIP
Sample
Human Cell lysate - whole cell (Uterus)
Specification
Uterus
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Detection step
Semiquantitative PCR
Positive control
RNA Polymerase II
Negative control
Rabbit IgG

Abcam user community

Verified customer

Submitted Oct 17 2008

Answer

Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab32085 in immunohistochemistry. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered. I have looked at the protocols you used and have a few questions and suggestions that might help you determine the cause of the no-staining results. I would therefore appreciate if you can please clarify the following items. - are your kidney samples taken from human? - can you confirm that the second antibody you used also managed to produce good results with other primary antibodies? Was the secondary antibody biotin conjugated? In some cases, the problem may be with the secondary antibody not working. - was the deparaffinization process complete? Did you used a new batch of xylene? - how long was the heating in citrate buffer done? We would normally recommend using 0.01M Citrate Buffer, pH6.0 and heat for 10 min. - After making the ABC complex, how long did you leave it to stand? In general, it will require a minimum of 30 minutes as this is the length of time that the complex takes to form. Did this ABC kit also work well in other recent experiments? I hope the above recommendations may already help you. If you have already tried the above suggestions and still experience problems, please do not hesitate to contact me with your shipping address/purchasing agent information. Also please advice me on how you would like to proceed with your enquiry, so that I can immediately arrange for a replacement or refund to you.

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