Profile of Human Disialoganglioside on M21 human melanoma cells. M21 cells were stained with either Purified Mouse anti-Human Disialoganglioside GD2 (Cat. No. 554272; Solid histogram) or a Purified Mouse IgG2a, κ Isotype Control (Cat. No. 556651; Open histogram), followed by a FITC-conjugated second step. Flow cytometry was performed on a BD FACScan™ flow cytometry system (BDIS, San Jose, CA).
Flow cytometric analysis of purified anti-human disialoganglioside GD2 on human mesenchymal stem cells (MSCs). Human mesenchymal stem cells (Lonza, Cat. No. PT-2501) were harvested and stained with Purified Mouse anti-Human Disialoganglioside GD2 (solid line) or a Purified Mouse IgG2a, κ Isotype Control (Cat. No. 556651; dashed line). The second step reagent was PE Goat anti-Mouse Ig (Multiple Adsorption; Cat. No. 550589). Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Profile of Human Disialoganglioside on M21 human melanoma cells. M21 cells were stained with either Purified Mouse anti-Human Disialoganglioside GD2 (Cat. No. 554272; Solid histogram) or a Purified Mouse IgG2a, κ Isotype Control (Cat. No. 556651; Open histogram), followed by a FITC-conjugated second step. Flow cytometry was performed on a BD FACScan™ flow cytometry system (BDIS, San Jose, CA).
Flow cytometric analysis of purified anti-human disialoganglioside GD2 on human mesenchymal stem cells (MSCs). Human mesenchymal stem cells (Lonza, Cat. No. PT-2501) were harvested and stained with Purified Mouse anti-Human Disialoganglioside GD2 (solid line) or a Purified Mouse IgG2a, κ Isotype Control (Cat. No. 556651; dashed line). The second step reagent was PE Goat anti-Mouse Ig (Multiple Adsorption; Cat. No. 550589). Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Product Details
BD Pharmingen™
GD2; Disialoganglioside GD2; Ganglioside GD2
Human (QC Testing), Mouse (Tested in Development)
Mouse BALB/c IgG2a
LAN-1 human neuroblastoma cells
Flow cytometry (Routinely Tested), Cytotoxicity, Immunofluorescence, Immunohistochemistry-frozen (Reported)
0.5 mg/ml
AB_395336
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Recommended Assay Procedures
Human neuroblastoma and melanoma cells are suggested as positive controls. Clone 14.G2a has also been reported for antibody-dependent and complement-mediated cytotoxicity of GD2 positive tumor cells and inhibit the growth of GD2 positive cells in athymic nude mice.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
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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.
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Format Details
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
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Citations & References
Development References (9)
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Cheresh DA, Honsik CJ, Staffileno LK, Jung G, Reisfeld RA. Disialoganglioside GD3 on human melanoma serves as a relevant target antigen for monoclonal antibody-mediated tumor cytolysis. Proc Natl Acad Sci U S A. 1985; 82(15):5155-5159. (Biology). View Reference
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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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Cheresh DA, Pierschbacher MD, Herzig MA, Mujoo K. Disialogangliosides GD2 and GD3 are involved in the attachment of human melanoma and neuroblastoma cells to extracellular matrix proteins. J Cell Biol. 1986; 102(3):688-696. (Biology). View Reference
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Cheresh DA, Rosenberg J, Mujoo K, Hirschowitz L, Reisfeld RA. Biosynthesis and expression of the disialoganglioside GD2, a relevant target antigen on small cell lung carcinoma for monoclonal antibody-mediated cytolysis. Cancer Res. 1986; 46(10):5112-5118. (Clone-specific: Flow cytometry, Immunofluorescence, Immunohistochemistry). 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). 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: Flow cytometry). 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. (Clone-specific: Cytotoxicity, Inhibition). 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). 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.
