RORγ (t) Antibody, anti-human/mouse, REAfinity™

Name: RORγ (t) Antibody, anti-human/mouse, REAfinity™
Availability: InStock

RORγ (t) Antibody, anti-human/mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA278

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

ICFC, MICS, IF, IHC, MC

Alternative names:

RORC, NR1F3, RORG, RZR-GAMMA, RZRG, TOR, RORgamma, Thor

Intracellular flow cytometry applications forRORγ (t) Antibody, anti-human/mouse, REAfinity™

APC
PE
Vio B515

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Reported:

26942062,

Products used:

130-124-035, 130-123-840

Protocols:

Intracellular flow cytometry staining protocol (Transcription Factor Staining Buffer Set (1:50)

Description:

Mouse thymocytes were stained with CD4-FITC. Cells were then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were stained with Anti-RORγ (t) antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. The specificity of the conjugated antibodies is confirmed by blocking the binding to ligand, using pure unconjugated antibodies (control: left images). Cell debris were excluded from the analysis based on scatter signals.

MICS (MACSima Imaging Cyclic Staining) applications forRORγ (t) Antibody, anti-human/mouse, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-124-035, 130-123-840

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forRORγ (t) Antibody, anti-human/mouse, REAfinity™

pure

Conjugate:

pure

Products used:

130-108-059

Extended validation for RORγ (t) Antibody, anti-human/mouse, REAfinity™

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

Flow cytometric comparison of different clones for RORγ (t). Thymocytes derived from B57BL/6 mice were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Transcription Factor Staining Buffer Set followed by intracellular staining with RORγ (t) antibodies. As a control, RORγ (t) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for RORγ (t). Thymocytes derived from B57BL/6 mice were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Transcription Factor Staining Buffer Set followed by intracellular staining with RORγ (t) antibodies. As a control, RORγ (t) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for RORγ (t). Thymocytes derived from B57BL/6 mice were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Transcription Factor Staining Buffer Set followed by intracellular staining with RORγ (t) antibodies. As a control, RORγ (t) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for RORγ (t). Thymocytes derived from B57BL/6 mice were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Transcription Factor Staining Buffer Set followed by intracellular staining with RORγ (t) antibodies. As a control, RORγ (t) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for RORγ (t) Antibody, anti-human/mouse, REAfinity™

Overview

Clone REA278 recognizes the retinoid-related orphan receptor-γ (RORγ) antigen and the RORγ (t) isoform, which are ligand-dependent transcription factors that play roles in multiple physiological processes. RORγ is expressed with highest levels in several tissues like muscle, kidney, and liver, whereas RORγ (t) is expressed almost exclusively in immature CD4

⁺

CD8

⁺

double-positive thymocytes. RORγ is the possible nuclear receptor for hydroxycholesterols, the binding of which strongly promotes coactivator recruitment. It is essential for thymopoiesis and the development of several secondary lymphoid tissues, including lymph nodes. RORγ (t) negatively regulates expression of Fas ligand and IL-2 production in T cells. It also binds to the TEA promoter and may be part of the pre-TCR activation cascade leading to the maturation of α/β T cells and may participate in the regulation of DNA accessibility in the TCR-Jα locus.

Additional information: Clone REA278 displays negligible binding to Fc receptors.

Alternative names

RORC, NR1F3, RORG, RZR-GAMMA, RZRG, TOR, RORgamma, Thor

Detailed product information

Technical specifications

Clone	REA278
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (I), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human, mouse
Antigen	RORγ (t)
Alternative names of antigen	RORC, NR1F3, RORG, RZR-GAMMA, RZRG, TOR, RORgamma, Thor
Molecular mass of antigen [kDa]	58 (56)(antibody detects two isoforms, molecular weight in brackets is for RORγ (t))
Distribution of antigen	lymphocytes, thymocytes
Entrez Gene ID	6097
RRID	AB_2802014, AB_2811585, AB_2811561, AB_2889744, AB_2889743, AB_2653369

Resources for RORγ (t) Antibody, anti-human/mouse, REAfinity™

Documents and Protocols

Brochures/posters

REAfinity™ Antibodies for mass cytometry - Recombinantly engineered to increase reproducibility

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for RORγ (t) Antibody, anti-human/mouse, REAfinity™

Publications

He, Y. W. et al. (1998) RORγ t, a novel isoform of an orphan receptor, negatively regulates Fas ligand expression and IL-2 production in T cells. Immunity 9(6): 797-806
Villey, I. et al. (1999) RORγT, a thymus-specific isoform of the orphan nuclear receptor RORγ / TOR, is up-regulated by signaling through the pre-T cell receptor and binds to the TEA promoter. Eur. J. Immunol. 29(12): 4072-4080
Jin, L. et al. (2010) Structural basis for hydroxycholesterols as natural ligands of orphan nuclear receptor RORγ. Mol Endocrinol. 24(5): 923-929
Moretti, S. et al. (2017) A mast cell-ILC2-Tʜ9 pathway promotes lung inflammation in cystic fibrosis. Nat Commun. 8: 14017

Applications

Extended validation