Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [1]
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Validation data
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- Product number
- 46-9477-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IDO Monoclonal Antibody (eyedio), PerCP-eFluor™ 710, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- 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 causeapoptosis 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 (Product # 88-8824-00) 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 (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- eyedio
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references IL-10(-/-) Enhances DCs Immunity Against Chlamydia psittaci Infection via OX40L/NLRP3 and IDO/Treg Pathways.
Li Q, Li X, Quan H, Wang Y, Qu G, Shen Z, He C
Frontiers in immunology 2021;12:645653
Frontiers in immunology 2021;12:645653
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Normal human peripheral blood cells were stimulated overnight with LPS (Product # 00-4976-03) then surface stained with Anti-Human CD11c FITC (Product # 11-0116-42) followed by fixation and permeabilization with the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00). The cells were then intracellularly stained with Mouse IgG1 K Isotype Control PerCP-eFluor® 710 (Product # 46-4714-82) (left) or Anti-Human IDO PerCP-eFluor® 710 (right). Cells in the monocyte and lymphocyte gate were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 5 DCs maturation, T cell proliferation, and IDO expression in the co-culture system. (A) Flow cytometry detection of CD86 and MHC-II DC surface markers; mean fluorescence intensity (MFI) was calculated at 72 hpi. CD86 and MHC-II were analyzed by pre-gated CD11c + cells, CD11c was gated by isotype. CD86 and MHC-II surface markers were significantly upregulated in the co-culture system of CD4 + T cells and IL-10 -/- DCs. Compared to the DCs from the zDC-DTR group, CD86 and MHC-II were significantly higher in the DD group. (B) Measurement of CD4 + T cell proliferation via BrdU assay in the co-culture system of CD4 + T cells and DCs at 72 h. Higher CD4 + T cell proliferation was observed in the IL-10 -/- and anti-IL-10 groups than the zDC-DTR group ( P