Preparation And Storage
Recommended Assay Procedures
Applications include flow cytometry (1-2 µg/1x10^6 cells). Other applications not routinely tested at BD Biosciences Pharmingen include immunohistochemistry of frozen tissue sections, immunofluorescence microscopy of cultured cells and antibody-dependent and complement mediated cytotoxicity of GD3 positive cells.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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Gangliosides are sialic-acid bearing glycolipids expressed on the surface of all mammalian cells, and are likely involved in mediating cell-substratum interactions. They are important target antigens for antibody-mediated cytolysis of human melanoma and neuroblastoma cells. Human melanoma cells produce gangliosides GD2 and/or GD3 which are present in substratum-attached material, and may play a significant role in the melanoma metastatic phenotype. Ganglioside GD3 is a major surface marker on most human melanoma cells. MB3.6 has been used to localize GD3 in the plasma membrane and in focal adhesion plaques of human melanoma cells. Clone MB3.6 has also been shown to lyse GD3 positive human melanoma cells by both antibody-dependent and complement-mediated cytotoxicity, as well as inhibit the growth of human melanoma cells in athymic nude mice. Clone MB3.6 specifically reacts with human GD3 ganglioside. It does not cross-react with a variety of other gangliosides purified from melanoma or neuroblastoma cells. The FM9 human melanoma cell line was used as the immunogen.
Development References (6)
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Cheresh DA, Harper JR, Schulz G, Reisfeld RA. Localization of the gangliosides GD2 and GD3 in adhesion plaques and on the surface of human melanoma cells. Proc Natl Acad Sci U S A. 1984; 81(18):5767-5771. (Immunogen: Cytotoxicity, Immunofluorescence). View Reference
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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. (Clone-specific: Flow cytometry). 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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Hakomori S. Tumor-associated carbohydrate antigens. Annu Rev Immunol. 1984; 2:103-126. (Biology). 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. (Biology). View Reference
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