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Multicolor flow cytometric analysis of IDO1 expression on human peripheral blood mononuclear cells (PBMCs). PBMCs 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, then fixed and permeabilized using BD Cytofix/Cytoperm™ solution (Cat. No. 554722) and BD Perm/Wash™ buffer (Cat. No. 554723). The cells were then stained with BD Horizon™ BV421 Mouse Anti-Human CD11b (Cat. No. 562632) and Alexa Fluor™ 488 Mouse Anti-Human IDO1 antibody (Cat. No. 567853/567854). Bivariate pseudocolor density plots showing the correlated expression of IDO1 (or Ig Isotype control staining) versus CD11b were derived from gated events with the forward and side light-scatter characteristics of intact PBMCs. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
BD Pharmingen™ Alexa Fluor™ 488 Mouse Anti-Human IDO1
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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® CompBeads 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.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
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- Alexa Fluor™ is a trademark of Life Technologies Corporation.
Companion Products
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.
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
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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.