Description: The AFKJS-9 monoclonal antibody reacts with the mouse, human and rhesus monkey RORgamma(t) protein. RORgamma is a member of the retinoic acid-related orphan receptor (ROR) family, which also includes RORalpha and RORbeta. ROR family proteins are ligand-dependent transcription factors that play roles in multiple physiological processes. RORgamma is expressed in several tissues including liver, lung, muscle, heart and kidney. Furthermore, it was discovered that alternative transcription results in the expression of an isoform, RORgamma(t), which is expressed exclusively in cells of the lymphoid compartment, namely CD4+CD8+ "double-positive" thymocytes, Th17 cells of the periphery and lymphoid tissue inducer (LTi) cells of lymphoid organs.
The RORgamma(t) isoform differs from RORgamma by three unique amino acids at its amino terminus. Therefore, the AFKJS-9 antibody will react with both the RORgamma and RORgamma(t) isoforms.
Applications Reported: This AFKJS-9 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This AFKJS-9 antibody has been tested by intracellular staining followed by flow cytometric analysis of mouse thymocytes using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol B: One step protocol for intracellular (nuclear) proteins" located at www.thermofisher.com/flowprotocols. This may be used at less than or equal to 0.125 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser
ROR gamma, a NR1 Thyroid Hormone-Like receptor, has been shown to affect thymopoiesis, bone metabolism, T-cell apoptosis, and lymphoid organogenesis. ROR gamma has been shown to promote thymocyte survival by activating the expression of the antiapoptotic protein Bcl-x(L). ROR gamma is also required for the development of lymph nodes and Peyer's patches. It has been shown that ROR gamma(t), a thymus-specific isoform of ROR gamma from mouse, inhibits Fas ligand expression and cytokine secretion in immature thymocytes. ROR gamma binds as a monomer to response elements composed of a single core motif, GGTCA, preceded by a 6 bp AT-rich sequence. ROR gamma expression has been documented in mouse thymus, adipose, bone, skeletal muscle, liver, and kidney, and in human skeletal muscle. ESTs have been isolated from human tissue libraries, including cancerous head/neck, germ cell, and prostate, and normal colon, head/neck, lung, liver/spleen, and pancreas.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: MGC129539; Nuclear receptor ROR-gamma; Nuclear receptor RZR-gamma; Nuclear receptor subfamily 1 group F member 3; RAR-related orphan nuclear receptor variant 2; RAR-related orphan receptor C; RAR-related orphan receptor C, isoform a; retinoic acid-binding receptor gamma; retinoid-related orphan receptor gamma; Retinoid-related orphan receptor-gamma; ROR Gamma; RP11-98D18.11-001; Thymus orphan receptor; TOR
Gene Aliases: EGK_01272; IMD42; NR1F3; RORC; RORG; RORgamma; RZR-GAMMA; RZRG; Thor; TOR