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Flow cytometric analysis of CA IX expression on U-87 MG cells. Cells from the human U-87 MG (Glioblastoma, ATCC HTB-14™) cell line were stained with PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058; dashed line histogram) or PE Mouse Anti-Carbonic Anhydrase IX antibody (Cat. No. 568277/568278; solid line histogram) at 0.25 μg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing CA IX expression (or Ig Isotype control staining) was derived from gated events with the he forward and side-light scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ PE Mouse Anti-Human Carbonic Anhydrase IX
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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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
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The M75 monoclonal antibody specifically recognizes an epitope in the extracellular N-terminal proteoglycan-like region of Carbonic Anhydrase IX (CA IX). This zinc metalloenzyme is a transmembrane protein that is encoded by CA9 (Carbonic anhydrase 9). This enzyme is expressed in epithelial cells and carcinoma cell lines. It catalyzes the reversible hydration of carbon dioxide and plays a role in various biological processes including respiration, bone resorption, calcification, pH regulation, and cellular adhesion. Carbonic Anhydrase IX may be involved in controlling cellular proliferation and associated with cellular transformation. It is often expressed by tumor cells in response to hypoxia.
Development References (6)
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Mondal UK, Doroba K, Shabana AM, et al. PEG Linker Length Strongly Affects Tumor Cell Killing by PEGylated Carbonic Anhydrase Inhibitors in Hypoxic Carcinomas Expressing Carbonic Anhydrase IX. Int J Mol Sci. 2021; 22:3. (Clone-specific: Western blot). View Reference
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Opavský R, Pastoreková S, Zelník V, et al. Human MN/CA9 gene, a novel member of the carbonic anhydrase family: structure and exon to protein domain relationships.. Genomics. 1996; 33(3):480-7. (Biology). View Reference
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Pastorekova S, Gillies RJ. The role of carbonic anhydrase IX in cancer development: links to hypoxia, acidosis, and beyond.. Cancer Metastasis Rev. 2019; 38(1-2):65-77. (Biology). View Reference
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Pastoreková S, Závadová Z, Kostál M, Babusíková O, Závada J. A novel quasi-viral agent, MaTu, is a two-component system.. Virology. 1992; 187(2):620-6. (Immunogen: Flow cytometry, Immunofluorescence, Western blot). View Reference
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Zat'ovicová M, Tarábková K, Svastová E, et al. Monoclonal antibodies generated in carbonic anhydrase IX-deficient mice recognize different domains of tumour-associated hypoxia-induced carbonic anhydrase IX.. J Immunol Methods. 2003; 282(1-2):117-34. (Clone-specific: Immunofluorescence, Immunohistochemistry). View Reference
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Závada J, Závadová Z, Pastorek J, Biesová Z, Jezek J, Velek J. Human tumour-associated cell adhesion protein MN/CA IX: identification of M75 epitope and of the region mediating cell adhesion.. Br J Cancer. 2000; 82(11):1808-13. (Clone-specific: Blocking, ELISA, Flow cytometry, Functional assay, Western blot). View Reference
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