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Two-color flow cytometric analysis of IL-33R (ST2) expression by mouse splenocytes. BALB/c mouse splenic leucocytes were stained with BD Horizon™ Fixable Viability Stain 510 (Cat. No. 564406) followed by staining with Alexa Fluor® 488 Rat Anti-Mouse CD4 antibody (Cat. No. 557667) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Plot) or BD Horizon BV421 Rat Anti-Mouse IL-33R (ST2) antibody (Cat. No. 566309/566310; Right Plot). The cells were then fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with Alexa Fluor® 647 Rat Anti-Mouse Foxp3 (Cat. No. 560401/560402). Two-color flow cytometric contour plots showing the correlated expression of Foxp3 versus IL-33R (ST2) [or Ig Isotype control staining] were derived from CD4-positive gated events with the forward and side light-scatter characteristics of live cell-discriminated intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
BD Horizon™ BV421 Rat Anti-Mouse IL-33R (ST2)
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Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
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Companion Products
The U29-93 monoclonal antibody specifically binds to the mouse Interleukin-33 Receptor (IL-33 Receptor, or IL-33R) which is also known as ST2. The IL-33R exists in either a type I transmembrane or soluble glycoprotein form. These IL-33R forms are encoded by the Il1rl1 (Interleukin-1 receptor-like 1) gene which belongs to the IL-1 Receptor family within the Ig superfamily. The IL-33R is expressed by subsets of T cells, including Th2-like cells and some regulatory T cells, as well as some innate lymphocytes, eosinophils, basophils, and mast cells. The IL-33R (also known as the IL-33R alpha subunit, or IL-33Rα) binds IL-33 and complexes with the IL-1R Accessory Protein (IL1RAP) to form a functional signaling receptor complex that can induce the production of T helper type 2 (Th2) cytokines. The soluble IL-33R may function as a decoy receptor which can block the binding of IL-33 to the transmembrane IL-33R. The IL-33R plays roles in inflammation, immunity, and allergy.
Development References (3)
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Hayakawa H, Hayakawa M, Kume A, Tominaga S. Soluble ST2 blocks interleukin-33 signaling in allergic airway inflammation.. J Biol Chem. 2007; 282(36):26369-80. (Biology). View Reference
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Matta BM, Lott JM, Mathews LR, et al. IL-33 is an unconventional Alarmin that stimulates IL-2 secretion by dendritic cells to selectively expand IL-33R/ST2+ regulatory T cells.. J Immunol. 2014; 193(8):4010-20. (Biology). View Reference
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Schiering C, Krausgruber T, Chomka A, et al. The alarmin IL-33 promotes regulatory T-cell function in the intestine.. Nature. 2014; 513(7519):564-8. (Biology). View Reference
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