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Multiparameter flow cytometric analysis of IDO1 expression on human peripheral blood mononuclear cells (PBMCs). PBMCs were cultured in complete tissue culture medium without (Left Column) or with (Right Column) 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 Alexa Fluor® 488 Mouse Anti-Human CD11b (Cat. No. 557701 or 561687) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557732; Top Row) or Alexa Fluor® 647 Mouse Anti-Human IDO1 antibody (Cat. No. 566648; Bottom Row) at 0.06ug/test. Two-color flow cytometric pseudocolor 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 viable PBMCs. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Immunohistofluorescent and Immunohistochemical staining of IDO1 in human spleen. Following antigen retrieval with BD Pharmingen™ Retrievagen A buffer (Cat. No. 550524), the sections of formalin-fixed, paraffin-embedded tissue were stained as described. The IDO1-positive cells are localized to the white pulp near the central arteries. Original magnification, 20×. Top Image: Staining with Alexa Fluor® 647 Mouse Anti-Human IDO1 (Cat. No. 566648, pseudocolored red) and BD Horizon™ BV421 Mouse Anti-Human CD45 (Cat. No. 563879 or 563880, pseudocolored green). Photography was performed on a Molecular Devices standard epifluorescence microscope. Bottom Image: Overnight staining with either Purified Mouse IgG1 κ Isotype Control (Cat. No. 550878, not shown) or Purified Mouse Anti-Human IDO1 (available by custom order). A three-step staining procedure that employs Biotin Goat Anti-Mouse Ig polyclonal secondary antibody (Cat. No. 550337), Streptavidin HRP (Cat. No. 550946) and DAB (Cat. No. 550880) and Hematoxylin counterstaining were used.
BD Pharmingen™ Alexa Fluor® 647 Mouse Anti-Human IDO1
BD Pharmingen™ Alexa Fluor® 647 Mouse Anti-Human IDO1
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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
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
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