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PE Mouse Anti-Human Carbonic Anhydrase IX
PE Mouse Anti-Human Carbonic Anhydrase IX
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.
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.
Product Details
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BD Pharmingen™
CA-IX; CAIX; CA9; CAH9; Carbonic anhydrase 9
Human (QC Testing)
Mouse BALB/c IgG2b
MaTu-infected HeLa Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
568277 Rev. 2
Antibody Details
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M75

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.

568277 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
568277 Rev.2
Citations & References
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View product citations for antibody "568277" on CiteAb

Development References (6)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
View All (6) View Less
568277 Rev. 2

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

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.