Anti-Retinoic Acid Receptor alpha [Ralpha10] antibody (ab5423)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab5423 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
ICC 1/10 - 1/200.
ICC/IF 1/10 - 1/200.
WB 1/100 - 1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 50 kDa).Can be blocked with Retinoic Acid Receptor alpha peptide (ab4910).

Due to the extremely low levels of RAR expressed in native tissue it is recommented that a sensitive detection system is use.

Target

  • FunctionReceptor for retinoic acid. Retinoic acid receptors bind as heterodimers to their target response elements in response to their ligands, all-trans or 9-cis retinoic acid, and regulate gene expression in various biological processes. The RXR/RAR heterodimers bind to the retinoic acid response elements (RARE) composed of tandem 5'-AGGTCA-3' sites known as DR1-DR5. In the absence of ligand, the RXR-RAR heterodimers associate with a multiprotein complex containing transcription corepressors that induce histone acetylation, chromatin condensation and transcriptional suppression. On ligand binding, the corepressors dissociate from the receptors and associate with the coactivators leading to transcriptional activation. RARA plays an essential role in the regulation of retinoic acid-induced germ cell development during spermatogenesis. Has a role in the survival of early spermatocytes at the beginning prophase of meiosis. In Sertoli cells, may promote the survival and development of early meiotic prophase spermatocytes. In concert with RARG, required for skeletal growth, matrix homeostasis and growth plate function (By similarity). Regulates expression of target genes in a ligand-dependent manner by recruiting chromatin complexes containing MLL5. Mediates retinoic acid-induced granulopoiesis.
  • Involvement in diseaseNote=Chromosomal aberrations involving RARA are commonly found in acute promyelocytic leukemia. Translocation t(11;17)(q32;q21) with ZBTB16/PLZF; translocation t(15;17)(q21;q21) with PML; translocation t(5;17)(q32;q11) with NPM. The PML-RARA oncoprotein requires both the PML ring structure and coiled-coil domain for both interaction with UBE2I, nuclear microspeckle location and sumoylation. In addition, the coiled-coil domain functions in blocking RA-mediated transactivation and cell differentiation.
  • Sequence similaritiesBelongs to the nuclear hormone receptor family. NR1 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • DomainComposed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
  • Post-translational
    modifications
    Phosphorylated on serine and threonine residues. Phosphorylation does not change during cell cycle. Phosphorylation on Ser-77 is crucial for transcriptional activity (By similarity). Phosphorylation by AKT1 is required for the repressor activity but has no effect on DNA binding, protein stability nor subcellular localization. Phosporylated by PKA in vitro. This phosphorylation on Ser-219 and Ser-369 is critical for ligand binding, nuclear localization and transcriptional activity in response to FSH signaling.
    Sumoylated by SUMO2, mainly on Lys-399 which is also required for SENP6 binding. On all-trans retinoic acid (ATRA) binding, a confromational change may occur that allows sumoylation on two additional site, Lys-166 and Lys-171. Probably desumoylated by SENP6. Sumoylation levels determine nuclear localization and regulate ATRA-mediated transcriptional activity.
    Trimethylation enhances heterodimerization with RXRA and positively modulates the transcriptional activation.
    Ubiquitinated.
  • Cellular localizationNucleus. Cytoplasm. Nuclear localization depends on ligand binding, phosphorylation and sumoylation. Transloaction to the nucleus in the absence of ligand is dependent on activation of PKC and the downstream MAPK phosphorylation.
  • Target information above from: UniProt accession P10276 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • Acute Promelocytic Leukemia Breakpoint Cluster Region antibody
    • Acute Promelocytic Leukemia Breakpoint Cluster Region antibody
    • NR1B1 antibody
    • Nuclear mitotic apparatus protein retinoic acid receptor alpha fusion protein antibody
    • Nuclear receptor subfamily 1 group B member 1 antibody
    • Nucleophosmin retinoic acid receptor alpha fusion protein NPM RAR long form antibody
    • RAR A antibody
    • RAR alpha antibody
    • RAR alpha form antibody
    • RAR alpha form antibody
    • RAR antibody
    • RAR-alpha antibody
    • RARA antibody
    • RARA protein antibody
    • RARA/PML Fusion Gene antibody
    • RARA/PML Fusion Gene antibody
    • RARA_HUMAN antibody
    • RARalpha antibody
    • RARalpha1 antibody
    • Retinoic acid receptor alpha antibody
    • Retinoic acid receptor alpha polypeptide antibody
    see all

Anti-Retinoic Acid Receptor alpha [Ralpha10] antibody images

  • Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y cells labeling Retinoic Acid Receptor (green) with ab5423 at 1/100. F-Actin staining with Phalloidin (red) and nuclei with DAPI or Hoechst (blue). Cells were fixed with formalin, permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA in PBS for 30 minutes at room temperature. Cells were incubated with the primary antibody in 3% BSA in PBS overnight at 4°C. A DyLight-conjugated secondary antibody was used. 60X magnification. Left - negative control.

  • Immunocytochemistry/Immunofluorescence analysis of NIH-3T3 cells labeling Retinoic Acid Receptor (green) with ab5423 at 1/100. F-Actin staining with Phalloidin (red) and nuclei with DAPI or Hoechst (blue). Cells were fixed with formalin, permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA in PBS for 30 minutes at room temperature. Cells were incubated with the primary antibody in 3% BSA in PBS overnight at 4°C. A DyLight-conjugated secondary antibody was used. 60X magnification. Left - negative control.

  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labeling Retinoic Acid Receptor (green) with ab5423 at 1/100. F-Actin staining with Phalloidin (red) and nuclei with DAPI or Hoechst (blue). Cells were fixed with formalin, permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA in PBS for 30 minutes at room temperature. Cells were incubated with the primary antibody in 3% BSA in PBS overnight at 4°C. A DyLight-conjugated secondary antibody was used. 60X magnification. Left - negative control.

References for Anti-Retinoic Acid Receptor alpha [Ralpha10] antibody (ab5423)

ab5423 has not yet been referenced specifically in any publications.

Product Wall

Thank you for your reply. I have processed your request to receive ab28767 as a replacement for ab5423 which did not work as guaranteed in western blot. Your new order number is *******. If there is anything else I can help with, please let me know.

Thank you for contacting Abcam. I am unable to find the original protocol that was used to test ab5423. I do have some information on published references, however. The following protocol comes from this reference: Masarrat Ali, et al. (1992). Identifi...

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Application Immunocytochemistry/ Immunofluorescence
Sample Hamster Cell (CHO-K1 cells expressing Human Retinoic Acid Recept)
Specification CHO-K1 cells expressing Human Retinoic Acid Recept
Fixative Formaldehyde
Permeabilization Yes - 0.1 % Triton X-100
Blocking step TBS + 3% Dried Milk + 0.1 % Triton X-100 as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 22°C
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Dr. Mahesh Mathrubutham

Verified customer

Submitted Sep 14 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"