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Flow cytometric analysis using BD OptiBuild™ BV421 Mouse Anti-Human MIC A/B antibody (Cat. No. 742322; solid line histogram) on HeLa cells, with corresponding Isotype Control (dotted line histogram). Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
BD OptiBuild™ BV421 Mouse Anti-Human MIC A/B
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
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
Product Notices
- This antibody was developed for use in flow cytometry.
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
Companion Products
The 6D4 monoclonal antibody specifically binds to the human MHC class I polypeptide-related sequence A (MICA, aka PERB11.1) and B (MICB. aka PERB11.2) proteins. These ~70 kDa transmembrane glycoproteins are homologs of the major histocompatibility complex class I molecules although they lack association with β2 microglobulin. The MHC class I-related MICA and MICB chains are expressed by some gut epithelial cells in vivo. MICA and MICB expression by other epithelial cells and cell types, including fibroblasts and endothelial cells, is induced by stress, eg, stress caused by bacterial and viral infections, autoimmunity or cellular transformation. Epithelial cell expression of MICA and MICB has also been detected in transplanted kidneys and pancreas that show histological signs of rejection and or cellular injury. This suggests their potential role in transplant immunopathology. MICA and MICB are ligands for NKG2D (CD314), an activating receptor expressed by natural killer (NK) cells, γδ T cells, CD8+ and some CD4+ αβ T cells. The 6D4 antibody reportedly blocks NKG2D-positive NK cell- and T cell-mediated cytotoxicity against MICA/B-positive target cells.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (8)
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Bauer S, Groh V, Wu J, et al. Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science. 1999; 285(5428):727-729. (Clone-specific: Blocking). View Reference
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Das H, Groh V, Kuijl C, et al. MICA engagement by human Vgamma2Vdelta2 T cells enhances their antigen-dependent effector function. Immunity. 2001; 15(1):83-93. (Clone-specific: Blocking, Flow cytometry). View Reference
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Groh V, Bahram S, Bauer S, Herman A, Beauchamp M, Spies T. Cell stress-regulated human major histocompatibility complex class I gene expressed in gastrointestinal epithelium. Proc Natl Acad Sci U S A. 1996; 93(22):12445-12450. (Biology). View Reference
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Groh V, Bruhl A, El-Gabalawy H, Nelson JL, Spies T. Stimulation of T cell autoreactivity by anomalous expression of NKG2D and its MIC ligands in rheumatoid arthritis. Proc Natl Acad Sci U S A. 2003; 100(16):9452-9457. (Clone-specific: Blocking, Flow cytometry, Immunohistochemistry). View Reference
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Groh V, Rhinehart R, Randolph-Habecker J, Topp MS, Riddell SR, Spies T. Costimulation of CD8alphabeta T cells by NKG2D via engagement by MIC induced on virus-infected cells. Nat Immunol. 2001; 2(3):255-260. (Clone-specific: Blocking, Flow cytometry, Immunofluorescence, Immunohistochemistry). View Reference
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Groh V, Rhinehart R, Secrist H, Bauer S, Grabstein KH, Spies T. Broad tumor-associated expression and recognition by tumor-derived gamma delta T cells of MICA and MICB. Proc Natl Acad Sci U S A. 1999; 96(12):6879-6884. (Clone-specific: Blocking, Flow cytometry, Immunohistochemistry). View Reference
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Groh V, Steinle A, Bauer S, Spies T. Recognition of stress-induced MHC molecules by intestinal epithelial gammadelta T cells. Science. 1998; 279(5357):1737-1740. (Immunogen: Blocking, Flow cytometry). View Reference
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Hankey KG, Drachenberg CB, Papadimitriou JC, et al. MIC expression in renal and pancreatic allografts. Transplantation. 2002; 73(2):304-306. (Clone-specific: Immunohistochemistry). View Reference
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.