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BD Horizon™ BV421 Mouse Anti-Human Disialoganglioside GD2
Clone 14.G2a (also known as 14G2a) (RUO)
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.
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.
Product Details
BD Horizon™
GD2; Disialoganglioside GD2; Ganglioside GD2
Human (QC Testing)
Mouse BALB/c IgG2a
LAN-1 human neuroblastoma cells
Flow cytometry (Routinely Tested)
5 µl
AB_2738679
Aqueous buffered solution containing BSA and ≤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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
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
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
Brilliant Stain Buffer RUO
Size
1000 Tests
Cat No.
566349
BV421 Mouse IgG2a, k Isotype Control RUO
Size
50 µg
Cat No.
562439
Accutase™ Cell Detachment Solution RUO
Size
100 ML
Cat No.
561527
564223 Rev. 2
Antibody Details
14.G2a
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.
564223 Rev. 2
Format Details
BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
407 nm
423 nm
564223 Rev.2
Citations & References
View product citations for antibody "564223" on CiteAb
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
564223 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.