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Flow cytometric analysis of GD2 expression on human mesenchymal stem cells. Human bone marrow-derived mesenchymal stem cells (Lonza, Cat. No. PT-2501) at passage 5 were harvested using BD™ Accutase™ Cell Detachment Solution (Cat. No. 561527). The cells were washed and then stained with either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439, dashed line histogram) or BD Horizon BV421 Mouse anti-Human GD2 antibody (Cat. No. 564223/565991; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Flow cytometric analysis of GD2 expression on human M21 cells. Cells from the human M21 melanoma cell line were similarly stained with either BD Horizon BV421 Mouse IgG2a, κ Isotype Control (dashed line histogram) or BD Horizon BV421 Mouse Anti-Human GD2 antibody (solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BV421 Mouse Anti-Human Disialoganglioside GD2
BD Horizon™ BV421 Mouse Anti-Human Disialoganglioside GD2
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 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. 563794/566349).
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Accutase is a registered trademark of Innovative Cell Technologies, Inc.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Gangliosides are sialic-acid bearing glycolipids that are expressed on the surface of all mammalian cells, and are likely involved in mediating cell-substratum interactions. They are important target antigens for antibody dependent cellular cytotoxicity (ADCC) of human melanoma and neuroblastoma cells. Human melanoma cells produce gangliosides, designated as GD2 and GD3 which are deposited in the subtratum-attached material, and may play a significant role in the melanoma metastatic phenotype. Clone 14.G2a specifically reacts with human and mouse GD2 ganglioside. LAN-1 human neuroblastoma cells were used as immunogen. Clone 14.G2a is an isotype switch variant selected from the parental IgG3-producing hybridoma 14.18 and has identical reactivity as the parental antibody. Clone 14.G2a is routinely tested by flow cytometry using M21 human melanoma 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 (6)
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Cheresh DA, Klier FG. Disialoganglioside GD2 distributes preferentially into substrate-associated microprocesses on human melanoma cells during their attachment to fibronectin. J Cell Biol. 1986; 102(5):1887-1897. (Biology). View Reference
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Frost JD, Hank JA, Reaman GH, et al. A phase I/IB trial of murine monoclonal anti-GD2 antibody 14.G2a plus interleukin-2 in children with refractory neuroblastoma: a report of the Children's Cancer Group. Cancer. 1997; 80(2):317-333. (Clone-specific: Cytotoxicity, In vivo exacerbation). View Reference
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Hakomori S. Tumor-associated carbohydrate antigens. Annu Rev Immunol. 1984; 2:103-126. (Biology). View Reference
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Lode HN, Reisfeld RA, Handgretinger R, Nicolaou KC, Gaedicke G, Wrasidlo W. Targeted therapy with a novel enediyene antibiotic calicheamicin theta(I)1 effectively suppresses growth and dissemination of liver metastases in a syngeneic model of murine neuroblastoma. Cancer Res. 1998; 58(14):2925-2928. (Clone-specific: Cytotoxicity, In vivo exacerbation). View Reference
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Mujoo K, Cheresh DA, Yang HM, Reisfeld RA. Disialoganglioside GD2 on human neuroblastoma cells: target antigen for monoclonal antibody-mediated cytolysis and suppression of tumor growth. Cancer Res. 1987; 47(4):1098-1104. (Immunogen: ELISA, Flow cytometry, Immunohistochemistry). View Reference
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Mujoo K, Kipps TJ, Yang HM, et al. Functional properties and effect on growth suppression of human neuroblastoma tumors by isotype switch variants of monoclonal antiganglioside GD2 antibody 14.18. Cancer Res. 1989; 49(11):2857-2861. (Clone-specific: Cytotoxicity, In vivo exacerbation). View Reference
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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.