High-throughput imaging method for direct assessment of GM1 ganglioside levels in mammalian cells
GM1-gangliosidosis is an inherited autosomal recessive disorder caused by mutations in the gene GLB1, which encodes acid β-galactosidase (β-gal). The lack of activity in this lysosomal enzyme leads to accumulation of GM1 gangliosides (GM1) in cells. We have developed a high-content-imaging method to...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2016-03-01
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| Series: | Data in Brief |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352340916000330 |
| _version_ | 1811213310898470912 |
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| author | Walter Acosta Reid Martin David N. Radin Carole L. Cramer |
| author_facet | Walter Acosta Reid Martin David N. Radin Carole L. Cramer |
| author_sort | Walter Acosta |
| collection | DOAJ |
| description | GM1-gangliosidosis is an inherited autosomal recessive disorder caused by mutations in the gene GLB1, which encodes acid β-galactosidase (β-gal). The lack of activity in this lysosomal enzyme leads to accumulation of GM1 gangliosides (GM1) in cells. We have developed a high-content-imaging method to assess GM1 levels in fibroblasts that can be used to evaluate substrate reduction in treated GLB1−/− cells [1]. This assay allows fluorescent quantification in a multi-well system which generates unbiased and statistically significant data. Fluorescently labeled Cholera Toxin B subunit (CTXB), which specifically binds to GM1 gangliosides, was used to detect in situ GM1 levels in a fixed monolayer of fibroblasts. This sensitive, rapid, and inexpensive method facilitates in vitro drug screening in a format that allows a high number of replicates using low working volumes. Keywords: High-throughput imaging, GM1-gangliosidosis, Acid β-galactosidase, Cholera toxin B subunit |
| first_indexed | 2024-04-12T05:44:41Z |
| format | Article |
| id | doaj.art-d75c718fd7f54bd2b1448cb96c7145e3 |
| institution | Directory Open Access Journal |
| issn | 2352-3409 |
| language | English |
| last_indexed | 2024-04-12T05:44:41Z |
| publishDate | 2016-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Data in Brief |
| spelling | doaj.art-d75c718fd7f54bd2b1448cb96c7145e32022-12-22T03:45:31ZengElsevierData in Brief2352-34092016-03-01610161022High-throughput imaging method for direct assessment of GM1 ganglioside levels in mammalian cellsWalter Acosta0Reid Martin1David N. Radin2Carole L. Cramer3BioStrategies LC, State University, P.O. Box 2428, AR 72467, USABioStrategies LC, State University, P.O. Box 2428, AR 72467, USA; Arkansas Biosciences Institute & Department Biological Sciences, Arkansas State University, P.O. Box 639, Jonesboro, AR 72467, USABioStrategies LC, State University, P.O. Box 2428, AR 72467, USABioStrategies LC, State University, P.O. Box 2428, AR 72467, USA; Arkansas Biosciences Institute & Department Biological Sciences, Arkansas State University, P.O. Box 639, Jonesboro, AR 72467, USAGM1-gangliosidosis is an inherited autosomal recessive disorder caused by mutations in the gene GLB1, which encodes acid β-galactosidase (β-gal). The lack of activity in this lysosomal enzyme leads to accumulation of GM1 gangliosides (GM1) in cells. We have developed a high-content-imaging method to assess GM1 levels in fibroblasts that can be used to evaluate substrate reduction in treated GLB1−/− cells [1]. This assay allows fluorescent quantification in a multi-well system which generates unbiased and statistically significant data. Fluorescently labeled Cholera Toxin B subunit (CTXB), which specifically binds to GM1 gangliosides, was used to detect in situ GM1 levels in a fixed monolayer of fibroblasts. This sensitive, rapid, and inexpensive method facilitates in vitro drug screening in a format that allows a high number of replicates using low working volumes. Keywords: High-throughput imaging, GM1-gangliosidosis, Acid β-galactosidase, Cholera toxin B subunithttp://www.sciencedirect.com/science/article/pii/S2352340916000330 |
| spellingShingle | Walter Acosta Reid Martin David N. Radin Carole L. Cramer High-throughput imaging method for direct assessment of GM1 ganglioside levels in mammalian cells Data in Brief |
| title | High-throughput imaging method for direct assessment of GM1 ganglioside levels in mammalian cells |
| title_full | High-throughput imaging method for direct assessment of GM1 ganglioside levels in mammalian cells |
| title_fullStr | High-throughput imaging method for direct assessment of GM1 ganglioside levels in mammalian cells |
| title_full_unstemmed | High-throughput imaging method for direct assessment of GM1 ganglioside levels in mammalian cells |
| title_short | High-throughput imaging method for direct assessment of GM1 ganglioside levels in mammalian cells |
| title_sort | high throughput imaging method for direct assessment of gm1 ganglioside levels in mammalian cells |
| url | http://www.sciencedirect.com/science/article/pii/S2352340916000330 |
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