Multicolor flow cytometric analysis of IDO1 expression on human peripheral blood mononuclear cells (PBMC). PBMC were cultured in complete tissue culture medium without (Left Plot) or with (Right Plot) lipopolysaccharide (LPS) (1 ug/ml, 20-24h, 37°C). The cells were harvested, fixed with BD Cytofix™ Fixation Buffer (Cat. 554655) and washed and permeabilized using BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with PE Mouse Anti-Human CD11b (Cat. No. 561001) and BD Horizon™ R718 Mouse Anti-Human IDO1 (Cat. No. 568016/568017) antibodies. The bivariate pseudocolor density plots showing the correlated expression of IDO1 versus CD11b were derived from gated events with the forward and side light-scatter characteristics of intact PBMC. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Cell Analyzer and FlowJo™ software.
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Multicolor flow cytometric analysis of IDO1 expression on human peripheral blood mononuclear cells (PBMC). PBMC were cultured in complete tissue culture medium without (Left Plot) or with (Right Plot) lipopolysaccharide (LPS) (1 ug/ml, 20-24h, 37°C). The cells were harvested, fixed with BD Cytofix™ Fixation Buffer (Cat. 554655) and washed and permeabilized using BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with PE Mouse Anti-Human CD11b (Cat. No. 561001) and BD Horizon™ R718 Mouse Anti-Human IDO1 (Cat. No. 568016/568017) antibodies. The bivariate pseudocolor density plots showing the correlated expression of IDO1 versus CD11b were derived from gated events with the forward and side light-scatter characteristics of intact PBMC. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Cell Analyzer and FlowJo™ software.
Product Details
BD Horizon™
IDO; IDO-1; INDO; indoleamine 2,3-dioxygenase 1
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human IDO1 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Brilliant Stain Buffer Plus RUO
Size
1000 Tests
Cat No.
566385
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
PE Mouse Anti-Human CD11b RUO
Size
25 Tests
Cat No.
561001
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
568016 Rev. 1
Antibody Details
V50-1886
The V50-1886 monoclonal antibody specifically binds to human IDO1, or indoleamine 2, 3-dioxygenase 1, a rate-limiting enzyme in the catabolism of tryptophan. IDO1 is expressed in normal epithelial, endothelial and myeloid cells and in tumor cells. Its expression is induced by interferon gamma and increases in viral and bacterial infections. The reduction of tryptophan and increase in its metabolites that are mediated by IDO1 have critical immunosuppressive and bactericidal effects.
568016 Rev. 1
Format Details
R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
R718
Red 627-640 nm
695 nm
718 nm
568016 Rev.1
Citations & References
View product citations for antibody "568016" on CiteAb
Development References (6)
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Liu H, Shen Z, Wang Z, et al. Increased expression of IDO associates with poor postoperative clinical outcome of patients with gastric adenocarcinoma.. Sci Rep. 2016; 6:21319. (Biology). View Reference
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Nakamura T1, Shima T, Saeki A, et al. Expression of indoleamine 2, 3-dioxygenase and the recruitment of Foxp3-expressing regulatory T cells in the development and progression of uterine cervical cancer.. 2007; 98(6):874-881. (Biology). View Reference
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Takikawa O1, Kuroiwa T, Yamazaki F, Kido R.. Mechanism of interferon-gamma action. Characterization of indoleamine 2,3-dioxygenase in cultured human cells induced by interferon-gamma and evaluation of the enzyme-mediated tryptophan degradation in its anticellular activity.. J Biol Chem. 1988; 263(4):2041-2048. (Biology). View Reference
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Théate I, van Baren N, Pilotte L, et al. Extensive profiling of the expression of the indoleamine 2,3-dioxygenase 1 protein in normal and tumoral human tissues.. Cancer Immunol Res. 2015; 3(2):161-72. (Biology). View Reference
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Zamorina SA, Timganova VP, Bochkova MS, Khramtsov PV, Raev MB. Effect of pregnancy-specific β1-glycoprotein on indoleamine-2,3-dioxygenase activity in human monocytes.. Dokl Biol Sci. 2016; 469(1):206-8. (Biology). View Reference
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van Baren N, Van den Eynde BJ. Tryptophan-degrading enzymes in tumoral immune resistance.. Front Immunol. 2015; 6:34. (Biology). View Reference
568016 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.