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DAMP, an acidotropic pH indicator, can be used as a tool to visualize non-esterified cholesterol in cells.

Acta biochimica et biophysica Sinica (2015-01-15)
Weimin Li, Rong Wang, Shaojuan Zhang, Xu Li
ABSTRACT

Cholesterol-rich regions are attractive targets for studying metabolic disorders that involve accumulation of cholesterol. Despite efforts to develop probes for labelling cholesterol-rich regions in cells, few of these reagents have a low molecular weight. Previous studies have shown that the acidotropic pH indicator, N-{3-[(2,4-dinitrophenyl)amino]propyl}-N-(3-aminopropyl)methylamine dihydrochloride (DAMP), reacts with cholesterol-rich organelles, such as endocrine secretary granules from endocrine cells. In this study, we demonstrated that DAMP could react with free cholesterol in a dose-dependent manner, and DAMP was able to detect cholesterol-rich subcellular organelles. DAMP was sufficiently potent to detect free cholesterol-enriched organs, but was unable to detect atherosclerotic plaques primarily composed of esterified cholesterol. Taken together, these results demonstrate that DAMP facilitates the study of cholesterol-enriched lipid rafts and disorders which involve cholesterol accumulation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Cholesterol, Sigma Grade, ≥99%
Sigma-Aldrich
Cholesterol, powder, BioReagent, suitable for cell culture, ≥99%
Sigma-Aldrich
2,4-Dinitrophenol, moistened with water, ≥98.0%
Sigma-Aldrich
SyntheChol® NS0 Supplement, 500 ×, synthetic cholesterol, animal component-free, sterile-filtered, aqueous solution, suitable for cell culture
SAFC
Cholesterol, Plant-Derived
Sigma-Aldrich
Nitrogen, ≥99.998%
Sigma-Aldrich
Cholesterol, from sheep wool, ≥92.5% (GC), powder
Supelco
Cholesterol, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Cholesterol solution, certified reference material, 10 mg/mL in chloroform
Supelco
2,4-Dinitrophenol, PESTANAL®, analytical standard
Sigma-Aldrich
Sandoz 58-035, >98% (HPLC), powder
SAFC
Cholesterol, from sheep wool, Controlled origin, meets USP/NF testing specifications