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Multicolor flow cytometric analysis of IL-33R (ST2) expression on Mouse splenocytes. BALB/c mouse splenic leucocytes were stained with BD Horizon™ Fixable Viability Stain 510 (Cat. No. 564406) followed by staining with FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553047) and with either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Plot) or BD Horizon™ PE-CF594 Rat Anti-Mouse IL-33R (ST2) antibody (Cat. No. 567521; Right Plot) at 1.0 μg/test. The cells were then fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with BD Horizon™ BV421 Rat Anti-Mouse Foxp3 (Cat. No. 562996). The bivariate pseudocolor density plot showing the correlated expression of Foxp3 versus IL-33R (ST2) [or Ig Isotype control staining] was derived from CD4-positive gated events with the forward and side light-scatter characteristics of live cell-discriminated intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ PE-CF594 Rat Anti-Mouse IL-33R (ST2)
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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.
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
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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 (6)
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Cottagiri M, Nyandjo M, Stephens M, et al. In drug-induced, immune-mediated hepatitis, interleukin-33 reduces hepatitis and improves survival independently and as a consequence of FoxP3+ T-cell activity.. Cell Mol Immunol. 2019; 16(8):706-717. (Clone-specific: Flow cytometry). View Reference
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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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Li A, Herbst RH, Canner D, et al. IL-33 Signaling Alters Regulatory T Cell Diversity in Support of Tumor Development.. Cell Rep. 2019; 29(10):2998-3008.e8. (Clone-specific: Flow cytometry). 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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Oldenhove G, Boucquey E, Taquin A, et al. PD-1 Is Involved in the Dysregulation of Type 2 Innate Lymphoid Cells in a Murine Model of Obesity.. Cell Rep. 2018; 25(8):2053-2060.e4. (Clone-specific: Flow cytometry). 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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For Research Use Only. Not for use in diagnostic or therapeutic procedures.