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Purified anti-RORγt Antibody

Pricing & Availability
Clone
A21005D (See other available formats)
Regulatory Status
RUO
Other Names
retinoic acid receptor (RAR)-related orphan receptor gamma T, ROR gamma2, RAR Related Orphan Receptor C, TOR, Retinoic Acid-Binding Receptor Gamma, IMD42, NR1f3
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
a. A21005D_PURE_RORgammat_Antibody-110822
Whole cell extracts (15 µg total protein) from SR cells, Jurkat cells, mouse thymus, and mouse spleen were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 μg/mL of purified anti-RORγt (clone A21005D) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 643807) was used as a loading control at a 1:10,000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • a. A21005D_PURE_RORgammat_Antibody-110822
    Whole cell extracts (15 µg total protein) from SR cells, Jurkat cells, mouse thymus, and mouse spleen were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 μg/mL of purified anti-RORγt (clone A21005D) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 643807) was used as a loading control at a 1:10,000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • B. A21005D_PURE_RORgammat_Antibody-3_102522
    SR cells (positive control, filled histogram) and Jurkat cells (negative control, open histogram) were fixed and permeabilized using True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) and intracellularly stained with purified anti-RORγt (clone A21005D) or purified mouse IgG1, κ isotype control (Cat. No. 401402) (open histogram, dashed line) followed by PE goat anti-mouse IgG (Cat. No. 405307).
  • C. A21005D_PURE_RORgammat_Antibody-2_102522
    ICC staining of purified anti-RORγt (clone A21005D) on SR cells (positive control, left panel) and Jurkat cells (negative control, right panel). The cells were fixed with 4% PFA Fixation Buffer (Cat. No. 420801) and permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. The cells were then stained with 5.0 µg/mL of purified anti-RORγt (clone A21005D), followed by incubation with 2.5 µg/mL of Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405326) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801), and the slides were mounted with ProLong™ Gold Antifade Mountant. The image was captured with a 60X objective. Scale bar: 50 µm
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609051 25 µg $118
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609052 100 µg $347
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Description

RORγt is a member of the nuclear hormone receptor family of transcription factors. RORγt is the isoform 2 of the RORC gene. It is produced by alternative splicing where the two 5’-exons are replaced by an alternative exon downstream in the gene, this splicing causes a shorter protein carrying three distinctive amino acids at its N-terminal end which are not present on RORγ. RORγt is mainly expressed on immature CD4+/CD8+ thymocytes and Th17 cells. It is important on lymphoid organogenesis in lymph nodes and Peyer’s patches but not on spleen. It plays an important role in promoting differentiation of pro-inflammatory Th17 cells by reducing apoptosis. 

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide from human RORγt
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICFC, ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 1.0 µg/mL. For immunocytochemistry, a concentration range of 2.5 - 5.0 μg/mL is recommended. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This antibody does not cross react with mouse RORγ or RORγt.

RRID
AB_2927967 (BioLegend Cat. No. 609051)
AB_2927967 (BioLegend Cat. No. 609052)

Antigen Details

Structure
RORγt is a 497 amino acid protein with a predicted molecular weight of 56 kD.
Distribution

Expressed on different subsets of T cells (mature and immature), on thymus and lymph nodes. Also, it has been found expressed on Treg cells in colon.

Function
Promotion of pro-inflammatory Th17 cells by downregulation of Fas ligand and IL2 which reduce apoptosis.
Ligand/Receptor
Ligand are cholesterol metabolites
Cell Type
Lymphocytes, Macrophages, NK cells, NKT cells, T cells, Th17, Tregs
Biology Area
Adaptive Immunity, Immuno-Oncology, Immunology, Innate Immunity, Transcription Factors
Molecular Family
Innate Immune Signaling
Antigen References
  1. Castro G, et al. 2017. PloS One. 12: e0181868.
  2. Villey I, et al. 1999. Eur J Immunol. 29:4072-80.
  3. Lamb D, et al. 2021. Respir Res. 22:158.
Gene ID
6097 View all products for this Gene ID
UniProt
View information about RORgammat on UniProt.org
Go To Top Version: 1    Revision Date: 10/25/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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