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Multiparameter flow cytometric analysis using BD OptiBuild™ RB744 Mouse Anti-Human CD203c antibody (Cat. No. 757659; Right Plot) on Human peripheral blood lymphoid cells, with corresponding IgG Isotype Control (Cat. No. 570519; Left Plot). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
Multiparameter flow cytometric analysis using BD OptiBuild™ RB744 Mouse Anti-Human CD203c antibody (Cat. No. 757659; Right Plot) on Human peripheral blood lymphoid cells, with corresponding IgG Isotype Control (Cat. No. 570519; Left Plot). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
BD OptiBuild™ RB744 Mouse Anti-Human CD203c
BD OptiBuild™ RB744 Mouse Anti-Human CD203c
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
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
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- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The NP4D6 monoclonal antibody specifically binds to CD203c. CD203c is a type II transmembrane glycoprotein that is a member of the E-NNP family of ectoenzymes. CD203c is also known as ectonucleotide pyrophosphatase/phosphodiesterase 3 (E-NPP3, ENPP3) due to its capacity to hydrolyze phosphodiester and phosphosulfate bonds in a variety of molecules including deoxynucleotides, nucleoside phosphates, and nicotinamide adenine dinucleotide. CD203c is otherwise known as Phosphodiesterase-1β (PD-1β, PD-I beta), B10, or gp130RB13-6. CD203c is expressed by basophils and mast cells. Basophils increase CD203c expression following activation with allergens or antibodies that crosslink cytophilic IgE. Thus, CD203c serves as a useful flow cytometric marker for basophil activation and the detection and analysis of type 1 allergic responses. IgE-receptor cross-linking results in CD203c upregulation and overexpression on neoplastic mast cells in cases of systemic mastocytosis.
Development References (8)
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Binder M, Fierlbeck G, King T, Valent P, Buhring HJ. Individual hymenoptera venom compounds induce upregulation of the basophil activation marker ectonucleotide pyrophosphatase/phosphodiesterase 3 (CD203c) in sensitized patients. Int Arch Allergy Immunol. 2002; 129(2):160-168. (Clone-specific: Flow cytometry). View Reference
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Buhring HJ, Streble A, Valent P. The basophil-specific ectoenzyme E-NPP3 (CD203c) as a marker for cell activation and allergy diagnosis. Int Arch Allergy Immunol. 2004; 133(4):317-329. (Clone-specific: Flow cytometry). View Reference
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Fureder W, Schernthaner GH, Ghannadan M, et al. Quantitative, phenotypic, and functional evaluation of basophils in myelodysplastic syndromes. Eur J Clin Invest. 2001; 31(10):894-901. (Biology). View Reference
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Ghannadan M, Hauswirth AW, Schernthaner GH, et al. Detection of novel CD antigens on the surface of human mast cells and basophils. Int Arch Allergy Immunol. 2002; 127(4):299-307. (Clone-specific: Immunofluorescence). View Reference
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Hauswirth AW, Natter S, Ghannadan M, et al. Recombinant allergens promote expression of CD203c on basophils in sensitized individuals. J Allergy Clin Immunol. 2002; 110(1):102-109. (Clone-specific: Flow cytometry). View Reference
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Hauswirth AW, Sonneck K, Florian S, et al. Interleukin-3 promotes the expression of E-NPP3/CD203C on human blood basophils in healthy subjects and in patients with birch pollen allergy. Int J Immunopathol Pharmacol. 2007; 20(2):267-278. (Biology). View Reference
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Hennersdorf F, Florian S, Jakob A, et al.. Identification of CD13, CD107a, and CD164 as novel basophil-activation markers and dissection of two response patterns in time kinetics of IgE-dependent upregulation. Cell Res. 2005; 15(5):325-335. (Biology). View Reference
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Iwamoto T, Yuta A, Tabata T, et al. Evaluation of basophil CD203c as a predictor of carboplatin-related hypersensitivity reaction in patients with gynecologic cancer. Biol Pharm Bull. 2012; 35(9):1487-1495. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.