Recombinant
RabMAb

Anti-ROR gamma antibody [EPR20006] (ab207082)

Overview

  • Product name
    Anti-ROR gamma antibody [EPR20006]
    See all ROR gamma primary antibodies
  • Description
    Rabbit monoclonal [EPR20006] to ROR gamma
  • Host species
    Rabbit
  • Specificity
    Our data suggests that this antibody is specific to ROR gamma isoform 2 which is known as ROR gammaT.
  • Tested applications
    Suitable for: Flow Cyt, IHC-P, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse ROR gamma aa 1-300. The exact sequence is proprietary.
    Database link: P51450

  • Positive control
    • WB: His-tagged mouse ROR gamma recombinant protein (aa 1-266); Mouse thymus lysate. IHC-P: Mouse thymus and spleen tissues. Flow Cyt: Mouse thymocytes. IP: Mouse thymus lysate.
  • General notes

    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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab207082 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 1/50.
IHC-P 1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

WB 1/2000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
IP 1/30.

Target

  • Function
    Nuclear receptor that binds DNA as a monomer to ROR response elements (RORE) containing a single core motif half-site 5'-AGGTCA-3' preceded by a short A-T-rich sequence. Key regulator of cellular differentiation, immunity, peripheral circadian rhythm as well as lipid, steroid, xenobiotics and glucose metabolism (PubMed:19381306, PubMed:19965867, PubMed:22789990, PubMed:26160376, PubMed:20203100). Considered to have intrinsic transcriptional activity, have some natural ligands like oxysterols that act as agonists (25-hydroxycholesterol) or inverse agonists (7-oxygenated sterols), enhancing or repressing the transcriptional activity, respectively (PubMed:19965867, PubMed:22789990). Recruits distinct combinations of cofactors to target gene regulatory regions to modulate their transcriptional expression, depending on the tissue, time and promoter contexts. Regulates the circadian expression of clock genes such as CRY1, ARNTL/BMAL1 and NR1D1 in peripheral tissues and in a tissue-selective manner. Competes with NR1D1 for binding to their shared DNA response element on some clock genes such as ARNTL/BMAL1, CRY1 and NR1D1 itself, resulting in NR1D1-mediated repression or RORC-mediated activation of the expression, leading to the circadian pattern of clock genes expression. Therefore influences the period length and stability of the clock. Involved in the regulation of the rhythmic expression of genes involved in glucose and lipid metabolism, including PLIN2 and AVPR1A (PubMed:19965867). Negative regulator of adipocyte differentiation through the regulation of early phase genes expression, such as MMP3. Controls adipogenesis as well as adipocyte size and modulates insulin sensitivity in obesity. In liver, has specific and redundant functions with RORA as positive or negative modulator of expression of genes encoding phase I and Phase II proteins involved in the metabolism of lipids, steroids and xenobiotics, such as SULT1E1. Also plays also a role in the regulation of hepatocyte glucose metabolism through the regulation of G6PC and PCK1 (PubMed:19965867). Regulates the rhythmic expression of PROX1 and promotes its nuclear localization (PubMed:19381306, PubMed:19965867, PubMed:22789990, PubMed:26160376, PubMed:20203100). Plays an indispensable role in the induction of IFN-gamma dependent anti-mycobacterial systemic immunity (PubMed:26160376).
    Isoform 2: Essential for thymopoiesis and the development of several secondary lymphoid tissues, including lymph nodes and Peyer's patches. Required for the generation of LTi (lymphoid tissue inducer) cells. Regulates thymocyte survival through DNA-binding on ROREs of target gene promoter regions and recruitment of coactivaros via the AF-2. Also plays a key role, downstream of IL6 and TGFB and synergistically with RORA, for lineage specification of uncommitted CD4(+) T-helper (T(H)) cells into T(H)17 cells, antagonizing the T(H)1 program. Probably regulates IL17 and IL17F expression on T(H) by binding to the essential enhancer conserved non-coding sequence 2 (CNS2) in the IL17-IL17F locus. May also play a role in the pre-TCR activation cascade leading to the maturation of alpha/beta T-cells and may participate in the regulation of DNA accessibility in the TCR-J(alpha) locus.
  • Tissue specificity
    Isoform 1 is widely expressed in many tissues, including liver and adipose, and highly expressed in skeletal muscle. Isoform 2 is primarily expressed in immature thymocytes.
  • Involvement in disease
    Immunodeficiency 42
  • Sequence similarities
    Belongs to the nuclear hormone receptor family. NR1 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • Domain
    The AF-2 (activation function-2) motif is required for recruiting coregulators containing LXXLL motifs such as NCOA1 and NCOA2.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • IMD42 antibody
    • MGC129539 antibody
    • NR1F3 antibody
    • Nuclear receptor ROR gamma antibody
    • Nuclear receptor ROR-gamma antibody
    • Nuclear receptor RZR gamma antibody
    • Nuclear receptor RZR-gamma antibody
    • Nuclear receptor subfamily 1 group F member 3 antibody
    • RAR related orphan nuclear receptor variant 2 antibody
    • RAR related orphan receptor C antibody
    • RAR related orphan receptor C, isoform a antibody
    • RAR related orphan receptor gamma antibody
    • RAR-related orphan receptor C antibody
    • Retinoic acid binding receptor gamma antibody
    • Retinoid related orphan receptor gamma antibody
    • Retinoid-related orphan receptor-gamma antibody
    • Rorc antibody
    • RORG antibody
    • RORG_HUMAN antibody
    • RZR GAMMA antibody
    • RZRG antibody
    • TOR antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling ROR gamma with ab207082 at 1/3000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on mouse thymus cortex, located in the left and right of the image, with sporadic nuclear staining on medulla, middle region (PMID:14691482).

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling ROR gamma with ab207082 at 1/3000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on the cortical region of mouse thymus shown at a high magnification (400×) (PMID:14691482).

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Lane 1 : Anti-ROR gamma antibody [EPR20006] (ab207082) at 1/10000 dilution
    Lanes 2-3 : Anti-ROR gamma antibody [EPR20006] (ab207082) at 1/1000 dilution

    Lane 1 : His-tagged mouse ROR gamma recombinant protein (aa 1-266)
    Lane 2 : His-tagged mouse ROR alpha recombinant protein (aa 48-215)
    Lane 3 : His-tagged mouse ROR beta recombinant protein (aa 9-140)

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 58 kDa
    Observed band size: 35 kDa (why is the actual band size different from the predicted?)



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 3 seconds; Lane 2/3: 3 minutes.

     

  • All lanes : Anti-ROR gamma antibody [EPR20006] (ab207082) at 1/2000 dilution

    Lane 1 : Mouse thymus lysate
    Lane 2 : Mouse liver lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 58 kDa
    Observed band size: 58 kDa


    Exposure time: 3 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile is consistent with the literature: PMID 14691482; PMID 9881970.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling ROR gamma with ab207082 at 1/3000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Sporadic nuclear staining on mouse spleen (PMID:14691482).

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed Mouse thymocytes labeling ROR gamma with ab207082 at 1/50 dilution. Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

    Note: Cell surface staining with PE conjugated anti-mouse CD8 and Alexa Fluor® 647 conjugated anti mouse CD4 antibodies, followed by fixation with 4% PFA for 10 min and intracellular staining with ab207082. The image shows thymocytes in the CD4+CD8+ (red) or the CD4+CD8- (black, negative control) gate. 

  • ROR gamma was immunoprecipitated from 0.35 mg of Mouse thymus lysate with ab207082 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207082 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

    Lane 1: Mouse thymus lysate, 10 μg (Input).

    Lane 2: ab207082 IP in Mouse thymus lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207082 inMouse thymus lysate.

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

    Exposure time: 10 seconds.

References

ab207082 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Ph9 and Ph6
Permeabilization
No
Specification
tonsil
Fixative
Formaldehyde
Username

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Verified customer

Submitted Jul 31 2018

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

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