1987
DOI: 10.1111/j.1432-1033.1987.tb10813.x
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Abstract: Gesellschaft fur Biotechnologische Forschung mbH BraunschweigMouse spleen cells were prepared from CBA/J mice, and T lymphocytes were selectively stimulated with the T cell mitogen concanavalin A and further propagated in the presence of the T cell growth factor interleukin-2. The T cells were metabolically labeled with ~-[l-'~C]galactose and D[1-'4C]glUcOSamine, and the gangliosides were extracted and purified by DEAE-Sepharose column chromatography. Carbohydrate backbone structures of the asialogangliosides,… Show more

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Cited by 84 publications
(50 citation statements)
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“…Further elongation to Gg 4 Cer is carried out by galactosyl-(␤1-3)-transferase, and sialylation of Gg 4 Cer by sialyltransferase-IV leads to the formation of GM1b, a ganglioside characteristic of mouse spleen (Figure 1; Nakamura et al 1987). GM1b is particularly expressed by mouse splenic T-lymphocytes (Figure 1; Müthing et al 1987) and thymocytes ( Figure 1; Müthing et al 1989). We were not able to detect GM1b ganglioside, because in thin-layer chromatography GM1b is recognized by anti-Gg 4 Cer antibody after Vibrio cholerae neuraminidase treatment (Müth-ing 1996).…”
Section: Discussionmentioning
confidence: 99%
“…Further elongation to Gg 4 Cer is carried out by galactosyl-(␤1-3)-transferase, and sialylation of Gg 4 Cer by sialyltransferase-IV leads to the formation of GM1b, a ganglioside characteristic of mouse spleen (Figure 1; Nakamura et al 1987). GM1b is particularly expressed by mouse splenic T-lymphocytes (Figure 1; Müthing et al 1987) and thymocytes ( Figure 1; Müthing et al 1989). We were not able to detect GM1b ganglioside, because in thin-layer chromatography GM1b is recognized by anti-Gg 4 Cer antibody after Vibrio cholerae neuraminidase treatment (Müth-ing 1996).…”
Section: Discussionmentioning
confidence: 99%
“…The supernatants were pooled, dried by rotary evaporation, and co-extracted phospholipids were removed by mild saponifi cation ( 62,63 ). Neutral GSLs were isolated by anion-exchange column chromatography using DEAE-Sepharose CL-6B (GE Healthcare, Munich, Germany) according to standard procedures ( 64,65 ) and fi nally dissolved in chloroform/methanol (2/1, v/v).…”
Section: Purifi Cation Of Neutral Gsls From Vero-b4 Cellsmentioning
confidence: 99%
“…Briefly, GSLs were extracted with chloroform/methanol (2/1, v/v), chloroform/methanol (1/1, v/v) and chloroform/methanol (1/2, v/v) (10-fold volumes of the tissue wet weight) for 30 min with sonication. The combined extracts were evaporated and resuspended in chloroform/methanol/water (30/60/8, by volume); gangliosides were isolated by anion exchange chromatography on DEAE-Sepharose CL-6B (Pharmacia, Freiburg, Germany) [20]. The ganglioside fraction was incubated for 1 h at 37°C in aqueous 1 N NaOH to saponify phospholipids, followed by neutralization with acetic acid and dialysis.…”
Section: Isolation Of Gsls From Tissuesmentioning
confidence: 99%