Glycosphingolipid immunostaining: Detection of antibody binding with an avidin-biotin enzyme system

Buehler, Joanne; Macher, Bruce A.

doi:10.1016/0003-2697(86)90551-8

Cited by 40 publications

(11 citation statements)

References 16 publications

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“…4). The authors prefer this strategy, but numerous immunoenzymatic methods such as the immunoperoxidase technique (Higashi et al, 1984) and the avidin-biotin enzyme system have been established as well (Buehler & Macher, 1986). A companion plate is chromatographed for comparison reasons in the same chamber under identical conditions and subjected to chemical detection (see above Section III.A).…”

Section: General Proceduresmentioning

confidence: 99%

Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry

Müthing

Distler

2009

Mass Spectrom. Rev.

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Glycosphingolipids (GSLs), composed of a hydrophilic carbohydrate chain and a lipophilic ceramide anchor, play pivotal roles in countless biological processes, including infectious diseases and the development of cancer. Knowledge of the number and sequence of monosaccharides and their anomeric configuration and linkage type, which make up the principal items of the glyco code of biologically active carbohydrate chains, is essential for exploring the function of GSLs. As part of the investigation of the vertebrate glycome, GSL analysis is undergoing rapid expansion owing to the application of novel biochemical and biophysical technologies. Mass spectrometry (MS) takes part in the network of collaborations to further unravel structural and functional aspects within the fascinating world of GSLs with the ultimate aim to better define their role in human health and disease. However, a single-method analytical MS technique without supporting tools is limited yielding only partial structural information. Because of its superior resolving power, robustness, and easy handling, high-performance thin-layer chromatography (TLC) is widely used as an invaluable tool in GSL analysis. The intention of this review is to give an insight into current advances obtained by coupling supplementary techniques such as TLC and mass spectrometry. A retrospective view of the development of this concept and the recent improvements by merging (1) TLC separation of GSLs, (2) their detection with oligosaccharide-specific proteins, and (3) in situ MS analysis of protein-detected GSLs directly on the TLC plate, are provided. The procedure works on a nanogram scale and was successfully applied to the identification of cancer-associated GSLs in several types of human tumors. The combination of these two supplementary techniques opens new doors by delivering specific structural information of trace quantities of GSLs with only limited investment in sample preparation.

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Section: General Proceduresmentioning

confidence: 99%

Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry

Müthing

Distler

2009

Mass Spectrom. Rev.

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“…This was done using the monoclonal antibody 01, provided by Dr. S.E. Pfeiffer (15), and the Avidin-Biotin alkaline phosphatase kit (Vector Laboratories, Burlingame, CA) (16). Paragloboside was detected in a similar way using the antibody F1Hll from Dainabot (Chiba, Japan) (17}.…”

Section: Methodsmentioning

confidence: 99%

Glycolipids of a human glioma cell line bearing receptors for platelet‐derived growth factor (PDGF)

Yates

Agudelo

Sung

1992

Lipids

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Glycolipids of U-1242 MG were characterized because results of previous studies showed that exogenous gangliosides, especially GM3, inhibit PDGF-stimulated growth of this human glioma cell line. GM3 and GM2 are the major gangliosides; both separate as doublets with thin-layer chromatography. The major neutral glycolipid is glucocerebroside with nonhydroxy fatty acids, but paragloboside, ceramide dihexoside, globoside, and asialoGM2 (GA2) are also present. The coexistence in U-1242 MG of these gangliosides and the PDGF receptor, whose mitogenic signal is modulated by GM3 in these cells, suggests a possible functional relationship among them with respect to growth regulation.

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“…HPTLC immunostaining with MoAbs was performed as described using an avidin-linked alkaline phosphatase detection method (Vector Laboratories, Burlingame, CA). 21,60 HPTLC immunostaining with B19 capsids and Stx was performed as described. 21,25 All experiments were performed in triplicate.…”

Section: Hptlc Immunostainingmentioning

confidence: 99%

Glycosphingolipid expression in acute nonlymphocytic leukemia: common expression of shiga toxin and parvovirus B19 receptors on early myeloblasts

Cooling

Zhang

Naides

et al. 2003

Blood

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Glycosphingolipids (GSLs) are complex macromolecules on cell membranes that have been shown to play a role in neutrophil differentiation, activation, phagocytosis, and adhesion to both microorganisms and vascular endothelium. Because GSLs are often cryptic antigens on cell membranes, little is known regarding GSL expression in early myelopoiesis. To study the latter, myeloblasts were collected from patients with acute nonlymphocytic leukemia (ANLL) who required therapeutic leukocytopheresis for hyperleukocytosis. The neutral GSLs were isolated and identified by high-performance thin-layer chromatography (HPTLC), HPTLC immunostaining, gas chromatography, nuclear magnetic resonance, and fast atom bombardment-mass spectrometry. Like mature peripheral blood neutrophils, myeloblasts expressed glucosylceramide, lactosylceramide, and the neolacto-family GSLs, lactotriaosylceramide and neolactotetraosylceramide. Unlike neutrophils and chronic myeloid leukemia, most ANLL samples also expressed the globo-series GSLs, globotriaosylceramide and globotetraosylceramide. Globo GSL expression was strongly associated with a myeloblastic (ANLL M0-M2) and monoblastic phenotype (M5). A weak association was also noted with expression of either lymphoid (P < .

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Glycosphingolipid immunostaining: Detection of antibody binding with an avidin-biotin enzyme system

Cited by 40 publications

References 16 publications

Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry

Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry

Glycolipids of a human glioma cell line bearing receptors for platelet‐derived growth factor (PDGF)

Glycosphingolipid expression in acute nonlymphocytic leukemia: common expression of shiga toxin and parvovirus B19 receptors on early myeloblasts

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