Disialoganglioside

HPTLC analysis by using eluted ganglioside was performed as previously described [35 (link)]. 10 µg/µL of purified gangliosides in chloroform/methanol (1:1, v/v) were spotted using a capillary tube on 10 × 10 cm TLC plates (Merck Millipore, MA, USA) and then developed with 100 mL chloroform/methanol/0.25% CaCl₂·H₂O (50:40:10, v/v/v). The developed gangliosides were stained with resorcinol solution [HCl (Samchun, Seoul, Korea), 0.1 M CuSO₄·5H₂O (Sigma-Aldrich, St. Louis, MO, USA), resorcinol (Sigma-Aldrich, St. Louis, MO, USA), distilled water], and the stained samples on the HPTLC plate were dried at 105 ℃ in a dry-oven for at least three hours. Monosialoganglioside (Matreya LLC, State College, PA, USA) and disialoganglioside (Matreya LLC, State College, PA, USA) mixtures were used as standard markers. The quantitation of ganglioside expression of HPTLC was used densitometry program (Beta 4.0.3 from Scion Image, Frederick, MD, USA).

High-performance thin-layer chromatography (HPTLC) analysis of the gangliosides was performed using a 10 × 10 cm HPTLC 5651 plate (Merck, Rahway, NJ, USA), as previously described [68 (link)]. Purified gangliosides (100 μg of protein/50 μL/lane) were separated using the HPTLC plates, which were subsequently developed using chloroform/MeOH/0.25% CaCl₂·H₂O (50:40:10, v/v/v). Gangliosides were visualized via 0.2% resorcinol staining and then the HPTLC plates were dried at 105 °C in a drying oven for at least 3 h. Monosialoganglioside (Matreya LLC, State College, PA, USA), disialoganglioside (Matreya LLC, State College, PA, USA), and rat brain gangliosides were used as markers for individual ganglioside species.