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™ BUV496 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 496-nm. BD Horizon BUV496 can be excited by the ultraviolet laser (355 nm) and detected with a 515/30 nm filter with a 450LP. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into the channel detecting BD Horizon V500 or BV510 (eg, 525/40-nm filter). However, the spillover can be corrected through compensation as with any other dye combination.