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 650 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
Companion Products
The 3D6 monoclonal antibody specifically binds to c-Met (MET), which is also known as Hepatocyte growth factor receptor (HGFR) or Scatter factor receptor (SF receptor). c-Met is a 190 kDa single-pass type I transmembrane glycoprotein that is posttranslationally cleaved into a disulfide-linked extracellular α-chain and a transmembrane β-chain. c-Met functions as a receptor tyrosine kinase (RTK) that is normally involved in the development, regeneration, and survival of cells and tissues. c-Met is expressed on a variety of cell types including stem cells and progenitor cells, hepatocytes, keratinocytes, epithelial cells, endothelial cells, and neurons. c-Met is autophosphorylated when bound by its ligand, Hepatocyte Growth Factor (HGF). This leads to further activation of downstream signaling pathways that induce multiple responses including cellular migration, proliferation, survival, and angiogenesis. Abnormal c-Met expression or activity has been associated with tumorigenesis. The 3D6 antibody can reportedly function as an agonist by binding to and activating human but not mouse c-Met.
The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
Development References (5)
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Kong-Beltran M, Seshagiri S, Zha J, et al. Somatic mutations lead to an oncogenic deletion of met in lung cancer.. Cancer Res. 2006; 66(1):283-9. (Clone-specific: Activation, Bioassay, Functional assay, In vivo exacerbation, Stimulation). View Reference
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Nguyen TH, Loux N, Dagher I, et al. Improved gene transfer selectivity to hepatocarcinoma cells by retrovirus vector displaying single-chain variable fragment antibody against c-Met.. Cancer Gene Ther. 2003; 10(11):840-9. (Clone-specific: Bioassay, Flow cytometry, Functional assay). View Reference
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Ohashi K, Marion PL, Nakai H, et al. Sustained survival of human hepatocytes in mice: A model for in vivo infection with human hepatitis B and hepatitis delta viruses.. Nat Med. 2000; 6(3):327-31. (Immunogen: Activation, Functional assay, In vivo exacerbation, Stimulation). View Reference
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Sakai K, Aoki S, Matsumoto K. Hepatocyte growth factor and Met in drug discovery.. J Biochem. 2015; 157(5):271-84. (Biology). View Reference
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Wright TG, Singh VK, Li JJ, et al. Increased production and secretion of HGF alpha-chain and an antagonistic HGF fragment in a human breast cancer progression model.. Int J Cancer. 2009; 125(5):1004-15. (Clone-specific: ELISA). 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.