IDO Monoclonal Antibody (eyedio), PerCP-eFluor™ 710, eBioscience™, Invitrogen™

Description

Description: This eyedio monoclonal antibody reacts with human indoleamine-2,3-dioxygenase (IDO, INDO, IDO1), an intracellular enzyme that catalyzes the degradation of tryptophan to kynurenines. IDO is expressed in a wide variety of tissues and cells, including macrophages, plasmacytoid dendritic cells, and several cell lines. IDO can be induced in many different cell types by IFN gamma or other inflammatory stimuli. Expression of IDO in antigen presenting cells and tumors is thought to mediate immune suppression through depletion of this essential amino acid and/or by the creation of tryptophan metabolites that cause apoptosis of T cells and induction of regulatory T cells. Applications Reported: This eyedio antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This eyedio antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of stimulated normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (cat. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This antibody can be used at 5 μL (0.06 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. 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 (cat. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) 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. Filtration: 0.2 μm post-manufacturing filtered. Gamma-interferon (IFNG, MIM 147570) has an antiproliferative effect on many tumor cells and inhibits intracellular pathogens such as Toxoplasma and Chlamydia, at least partly because of the induction of indoleamine 2,3-dioxygenase (INDO, EC 1.13.11.52). This enzyme catalyzes the degradation of the essential amino acid L-tryptophan to N-formyl-kynurenine.