Anti-ROR gamma T (RORC2, NR1F3, RORgammaT) (PE)

ROR-gamma T is a member of the orphan nuclear receptor family. ROR-gamma is expressed in the muscle, thymus, testis, pancreas, prostate, heart, and liver. ROR-gamma plays a role in thymocyte development and homeostasis. RORs bind to DNA as monomers on half-site elements with 5'A/T-rich extensions. An N-terminal isoform of ROR-gamma, RORgammaT, has been shown to be specifically expressed in the thymus. Applications: Suitable for use in Flow Cytometry. Other applications not tested. Recommended Dilutions: Flow Cytometry: 10ul labels < 1x10e6 cells. For intracellular staining, cells must first be fixed and permeabilized. 4% PFA is recommended as a fixative and a 0.1% saponin balanced salt solution is recommended for permeabilization and washing., Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: 1. Harvest cells and wash twice in saline buffer., 2. Cell surface staining may be done at this point following the manufacturer's staining procedure., 3. Resuspend up to 1x10e6 cells in 0.5ml of 4% paraformaldehyde fixation buffer and incubate at RT for 10 minutes., 4. Following fixation, wash the cells twice in saline buffer, then once in saponin-containing saline buffer., 5. After permeabilization, add 10ul of R1183-30H and incubate the cells for 30 minutes at RT in the dark., 6. Wash the cells twice with the saponin-containing saline buffer., 7. Resuspend the cells in saline buffer for final flow cytometric analysis. As a control, cells in a separate tube should be treated with PE-labeled mouse IgG2B antibody. Storage and Stability: Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: PE conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.