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RecombinantRabMAb

Recombinant Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)

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Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
  • Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
  • Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
  • Immunoprecipitation - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
  • Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
  • Immunoprecipitation - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
  • Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23871-271] to Retinoic Acid Receptor alpha - BSA and Azide free
  • Suitable for: IP, IHC-P, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free
    See all Retinoic Acid Receptor alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR23871-271] to Retinoic Acid Receptor alpha - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, IHC-P, WBmore details
    Unsuitable for: Flow Cyt or ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human testis tissue lysate; HeLa, Hep G2, NIH/3T3, PC-12, MCF7, U937 and HEK-203 whole cell lysates. IHC-P: Human testis tissue; Mouse testis tissue; Rat testis tissue. IP: HEK-293 and NIH/3T3 whole cell lysates.

  • General notes

    ab275748 is the carrier-free version of ab275745.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab275748 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
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
Use at an assay dependent concentration. Detects a band of approximately 52 kDa (predicted molecular weight: 51 kDa).
Notes
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
Use at an assay dependent concentration. Detects a band of approximately 52 kDa (predicted molecular weight: 51 kDa).
Application notes
Is unsuitable for Flow Cyt or ICC/IF.

Target

  • Function

    Receptor 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 disease

    Note=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 similarities

    Belongs to the nuclear hormone receptor family. NR1 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 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 localization

    Nucleus. 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.
  • Information by UniProt

  • Database links

    • Alternative names

      • 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 alpha antibody
      • RAR antibody
      • RAR-alpha antibody
      • rara antibody
      • RARA_HUMAN antibody
      • RARalpha antibody
      • RARalpha1 antibody
      • Retinoic acid nuclear receptor alpha variant 1 antibody
      • Retinoic acid nuclear receptor alpha variant 2 antibody
      • Retinoic acid receptor alpha antibody
      • Retinoic acid receptor alpha polypeptide antibody
      see all

    Images

    • Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      All lanes : Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] (ab275745) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : RARA CRISPR-Cas9 edited HeLa cell lysate
      Lane 3 : MCF7 cell lysate
      Lane 4 : HepG2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 51 kDa
      Observed band size: 40 kDa why is the actual band size different from the predicted?



      This data was developed using the same antibody clone in a different buffer formulation (ab275745).

      Lanes 1 - 4: Merged signal (red and green). Green - ab275745 observed at 40 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

      ab275745 was shown to react with Retinoic Acid Receptor alpha in wild-type HeLa cells in western blot. The bands observed in RARA knockout cell line ab265176 (RARA knockout cell lysate ab257629) below 40 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. HeLa wild-type and RARA knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab275745 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)

      This data was developed using ab275745, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Retinoic Acid Receptor alpha with ab275745 at 1/1000 (2.71 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on sertoli cells of human testis. The section was incubated with ab275745 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      All lanes : Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] (ab275745) at 1/1000 dilution

      Lane 1 : Human testis tissue lysate
      Lane 2 : Human brain tissue lysate
      Lane 3 : Human skeletal muscle tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

      Predicted band size: 51 kDa
      Observed band size: 52 kDa why is the actual band size different from the predicted?



      This data was developed using ab275745, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      Low expression: human brain, human skeletal muscle (Database:GTExPortal, HPA).

      Exposure time: 103 seconds.

    • Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      All lanes : Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] (ab275745) at 1/1000 dilution

      Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
      Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

      Predicted band size: 51 kDa
      Observed band size: 52 kDa why is the actual band size different from the predicted?



      This data was developed using ab275745, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      Exposure time: 70 seconds.

    • Immunoprecipitation - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      Immunoprecipitation - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)

      This data was developed using ab275745, the same antibody clone in a different buffer formulation.

      Retinoic Acid Receptor alpha was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate with ab275745 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275745 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10 ug

      Lane 2: ab275745 IP in HEK-293 whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275745 in HEK-293 whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 84 seconds.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)

      This data was developed using ab275745, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Retinoic Acid Receptor alpha with ab275745 at 1/1000 (0.542 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on sertoli cells of rat testis. The section was incubated with ab275745 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      Western blot - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      All lanes : Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] (ab275745) at 1/1000 dilution

      Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : U937 (human histiocytic lymphoma monocyte) whole cell lysate
      Lane 3 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
      Lane 4 : MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

      Predicted band size: 51 kDa



      This data was developed using ab275745, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: Low expression：MOLT-4.(Database:HPA)

      Lane 1: 8 seconds Lane2-4: 70 seconds

      Exposure time: 

    • Immunoprecipitation - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      Immunoprecipitation - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)

      This data was developed using ab275745, the same antibody clone in a different buffer formulation.

      Retinoic Acid Receptor alpha was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab275745 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275745 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug

      Lane 2: ab275745 IP in NIH/3T3 whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275745 in NIH/3T3 whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 84 seconds.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)

      This data was developed using ab275745, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Retinoic Acid Receptor alpha with ab275745 at 1/1000 (0.542 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on sertoli cells of mouse testis (PMID:16210368). The section was incubated with ab275745 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)
      Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] - BSA and Azide free (ab275748)

    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

    References (0)

    Publishing research using ab275748? Please let us know so that we can cite the reference in this datasheet.

    ab275748 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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