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    progesterone-receptor-antibody-yr85-ab32085.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families Zinc Finger
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RecombinantRabMAb

Recombinant Anti-Progesterone Receptor antibody [YR85] (ab32085)

  • Datasheet
  • SDS
Reviews (5)Q&A (4)References (25)

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Immunocytochemistry - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Immunocytochemistry - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Flow Cytometry (Intracellular) - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • OI-RD Scanning - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • 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

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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: 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.
  • 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
    For more information see here.

    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

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    YR85
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Zinc Finger
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • Nuclear Hormone Receptors
    • Progesterone
    • Neuroscience
    • Endocrine system
    • Gonadotrophic axis
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Progesterone
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • Nuclear hormone receptors
    • Progesterone
    • Cancer
    • Tumor biomarkers
    • Receptors
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-Progesterone Receptor antibody [YR85] - BSA and Azide free (ab206926)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • T-47D whole cell lysate (ab14899)
  • Recombinant Protein

    • Recombinant Human Progesterone Receptor protein (ab159105)

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
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Predicted molecular weight: 99 kDa.
IHC-P (4)
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.
Notes
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

  • 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.
  • Target information above from: UniProt accession P06401 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5241 Human
    • Omim: 607311 Human
    • SwissProt: P06401 Human
    • Unigene: 32405 Human
    • Unigene: 742403 Human
    • 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

    • Immunocytochemistry - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Immunocytochemistry - Anti-Progesterone Receptor antibody [YR85] (ab32085)

      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.

    • Immunocytochemistry - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Immunocytochemistry - Anti-Progesterone Receptor antibody [YR85] (ab32085)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progesterone Receptor antibody [YR85] (ab32085)

      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.

    • Flow Cytometry (Intracellular) - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Flow Cytometry (Intracellular) - Anti-Progesterone Receptor antibody [YR85] (ab32085)

      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. 

       

       

    • Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      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
    • Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      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 seconds


      Blocking buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)

      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.

    • OI-RD Scanning - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      OI-RD Scanning - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Anti-Progesterone Receptor antibody [YR85] (ab32085)

    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.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (25)

    Publishing research using ab32085? Please let us know so that we can cite the reference in this datasheet.

    ab32085 has been referenced in 25 publications.

    • 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
    • Cindrova-Davies T  et al. Menstrual flow as a non-invasive source of endometrial organoids. Commun Biol 4:651 (2021). PubMed: 34140633
    • Zhang X  et al. ID4 Promotes Breast Cancer Chemotherapy Resistance via CBF1-MRP1 Pathway. J Cancer 11:3846-3857 (2020). PubMed: 32328189
    • Kim YY  et al. Effects of Natural Progesterone and Synthetic Progestin on Germ Layer Gene Expression in a Human Embryoid Body Model. Int J Mol Sci 21:N/A (2020). PubMed: 31991577
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-5 of 5 Abreviews

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Progesterone Receptor antibody [YR85]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 27 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Progesterone Receptor antibody [YR85]

    Poor
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Mrs. Katie Elliott

    Verified customer

    Submitted Jul 15 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Progesterone Receptor antibody [Y85]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 26 2010

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Progesterone Receptor antibody [Y85]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 09 2009

    ChIP abreview for Anti-Progesterone Receptor antibody [Y85]

    Average
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 17 2008

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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