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RB705 Mouse Anti-Human Carbonic Anhydrase IX
Product Details
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BD OptiBuild™
CA-IX; CAIX; CA9; CAH9; Carbonic anhydrase 9
Human (Tested in Development)
Mouse BALB/c IgG2b
MaTu-infected HeLa Cells
Flow cytometry (Qualified)
0.2 mg/ml
768
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 that minimize unconjugated dye and antibody.

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

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  3. For U.S. patents that may apply, see bd.com/patents.
  4. 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.
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. 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.
  7. 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.
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. An isotype control should be used at the same concentration as the antibody of interest.
  11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  12. 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.
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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.

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Format Details
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RB705
The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
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RB705
Blue 488 nm
498 nm
707 nm
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Citations & References
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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
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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.