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RecombinantRabMAb

Recombinant Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)

Submit a review Submit a question References (1)
Western blot - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
  • Immunoprecipitation - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
  • Western blot - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
  • Immunoprecipitation - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23719-18] to ROR alpha/RORA
  • Suitable for: IP, IHC-P, WB
  • Reacts with: Mouse, Rat

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-ROR alpha/RORA antibody [EPR23719-18]
    See all ROR alpha/RORA primary antibodies

  • Description

    Rabbit monoclonal [EPR23719-18] to ROR alpha/RORA

  • Host species

    Rabbit

  • Specificity

    This antibody does not react with human species for immunoprecipitation application.

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Rat

  • Immunogen

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

  • Positive control

    • WB: Mouse and rat cerebellum tissue lysate. IHC-P: Mouse and rat cerebellum tissue. IP: Mouse and rat cerebellum tissue lysate.

  • General notes

    This product is produced using clone 18 of EPR23719.

    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 ab256799 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
1/30.
IHC-P
1/800. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Predicted molecular weight: 63 kDa.
Notes
IP
1/30.
IHC-P
1/800. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Predicted molecular weight: 63 kDa.
Application notes
Is unsuitable for ICC/IF or IHC-Fr.

Target

  • Function

    Orphan nuclear receptor. Binds DNA as a monomer to hormone response elements (HRE) containing a single core motif half-site preceded by a short A-T-rich sequence. This isomer binds to the consensus sequence 5'-[AT][TA]A[AT][CGT]TAGGTCA-3'. Regulates a number of genes involved in lipid metabolism such as apolipoproteins AI, APOA5, CIII, CYP71 and PPARgamma, in cerebellum and photoreceptor development including PCP2, OPN1SW, OPN1SM AND ARR3, in circadian rhythm with BMAL1, and skeletal muscle development with MYOD1. Possible receptor for cholesterol or one of its derivatives.

  • Tissue specificity

    Widely expressed in a number of tissues.

  • Sequence similarities

    Belongs to the nuclear hormone receptor family. NR1 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.

  • Post-translational
    modifications

    Phosphorylation by PKC in neurons inhibits transcriptional activity. Phosphorylated on Thr-216 by ERK2 in vitro.
    Sumoylated by SENP1 and SENP2. Sumoylation, promoted by PIAS2, PIAS3, PIASy but not PIAS1, enhances the transcriptional activity. Desumoylated by SENP1.
    Ubiquitinated. Ubiquitination is required for efficient transcriptional activity and is prevented by Hairless.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • fhl2a antibody
      • NR1F1 antibody
      • Nuclear receptor ROR alpha antibody
      • Nuclear receptor ROR-alpha antibody
      • Nuclear receptor RZR-alpha antibody
      • Nuclear receptor subfamily 1 group F member 1 antibody
      • RAR related orphan receptor A antibody
      • RAR related orphan receptor alpha antibody
      • RAR-related orphan receptor A antibody
      • Retinoid-related orphan receptor-alpha antibody
      • Rora antibody
      • RORA_HUMAN antibody
      • RZR alpha antibody
      • RZR-ALPHA antibody
      • RZRA antibody
      • Transcription factor RZR alpha antibody
      see all

    Images

    • Western blot - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
      Western blot - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
      All lanes : Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799) at 1/1000 dilution

      Lane 1 : Mouse cerebellum tissue lysate
      Lane 2 : Mouse spleen tissue 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: 63 kDa
      Observed band size: 60, 65 kDa why is the actual band size different from the predicted?



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

      Low expression: mouse spleen (PMID:9226375).

      The molecular weight  observed is consistent with what has been described in the literature (PMID:19955433, PMID:12023888)

       Exposure time: 3 minutes

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)

      Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling ROR alpha/RORA with ab256799 at 1/800 (0.601 µg/ml) dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse cerebellum (PMID:14966198). The section was incubated with ab256799 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunoprecipitation - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
      Immunoprecipitation - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)

      ROR alpha/RORA was immunoprecipitated from 0.35 mg mouse cerebellum tissue lysate with ab256799 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256799 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Mouse cerebellum tissue lysate 10µg

      Lane 2: ab256799 IP in mouse cerebellum tissue lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab256799 in mouse cerebellum tissue lysate

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

      Exposure time: 3 minutes.

    • Western blot - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
      Western blot - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
      Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799) at 1/1000 dilution + Rat cerebellum tissue lysate at 20 µg

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

      Predicted band size: 63 kDa
      Observed band size: 60, 65 kDa why is the actual band size different from the predicted?



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

      Exposure time: 3 minutes

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)

      Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling ROR alpha/RORA with ab256799 at 1/800 (0.601 µg/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat cerebellum. The section was incubated with ab256799 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunoprecipitation - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)
      Immunoprecipitation - Anti-ROR alpha/RORA antibody [EPR23719-18] (ab256799)

      ROR alpha/RORA was immunoprecipitated from 0.35 mg rat cerebellum tissue lysate with ab256799 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256799 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Rat cerebellum tissue lysate 10µg

      Lane 2: ab256799 IP in rat cerebellum tissue lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab256799 in rat cerebellum tissue lysate

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

      Exposure time: 3 minutes.

    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 (1)

    Publishing research using ab256799? Please let us know so that we can cite the reference in this datasheet.

    ab256799 has been referenced in 1 publication.

    • Li X  et al. MicroRNA-10a-3p Improves Cartilage Degeneration by Regulating CH25H-CYP7B1-RORa Mediated Cholesterol Metabolism in Knee Osteoarthritis Rats. Front Pharmacol 12:690181 (2021). PubMed: 34149433

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