Recombinant
RabMAb

Recombinant Anti-Progesterone Receptor antibody [SP2] (ab16661)

Rabbit monoclonal Progesterone Receptor antibody [SP2]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Rat, Human. Cited in 11 publication(s). Independently reviewed in 2 review(s).

Overview

  • Product name

    Anti-Progesterone Receptor antibody [SP2]
    See all Progesterone Receptor primary antibodies
  • Description

    Rabbit monoclonal [SP2] to Progesterone Receptor
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Rabbit
  • Immunogen

    Recombinant fragment within Human Progesterone Receptor aa 400-550. The exact sequence is proprietary.
    Database link: P06401

  • Positive control

    • Breast carcinomas
  • General notes

    This product was switched from a hybridoma to recombinant production method on 13th November 2019.

    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 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab16661 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
ICC/IF 1/100.
WB 1/200. Predicted molecular weight: 99,120 kDa.
IHC-P 1/400.

Staining of formalin-fixed tissues is required by boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.

Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

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

  • Immunocytochemistry/ Immunofluorescence analysis of T-47D (human ductal breast epithelial tumor epithelial cell) cells labeling Progesterone Receptor with purified ab16661 at 1/100 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow Cytometry analysis of T-47D (human ductal breast epithelial tumor epithelial cell) cells labeling Progesterone Receptor with purified ab16661 at 1/220 dilution (1.04 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue.

  • Immunohistochemistry analysis of human breast carcinoma tissue labelling SP2 with ab16661.

  • ab16661 staining rat ovary tissue sections by IHC-P.  Sections were fixed in 10% buffered formalin and subjected to heat mediated antigen retrieval in 10mM citrate buffer pH 6.0 prior to blocking with 5% serum for 20 minutes at 20°C.  The primary antibody was diluted 1/50 in TBS (with Tween) and incubated with the sample for 16 hours at 4°C.  A HRP conjugated goat anti-rabbit was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing T47D cells stained with ab16661 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16661, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:

  • Liu Y  et al. Efficacy and safety of TE/TEC/intensive paclitaxel neoadjuvant chemotherapy for the treatment of breast cancer. Oncol Lett 17:907-912 (2019). IHC-P, IHC ; Human . Read more (PubMed: 30655846) »
  • Sun L  et al. The Modified Bushen Antai Recipe Upregulates Estrogen and Progesterone Receptors at the Maternal-Fetal Interface in Pregnant Rats with Mifepristone-Induced Pregnancy Loss. Evid Based Complement Alternat Med 2019:8312020 (2019). IHC-P, IHC ; Rat . Read more (PubMed: 30792746) »
See all 24 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 20% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH9
Sample
Human Tissue sections (Breast carcinoma)
Specification
Breast carcinoma
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 15 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Rat ovary, uterus)
Specification
Rat ovary, uterus
Fixative
10% buffered formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citrate buffer pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 27 2007

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