title: Fluorescent Detection of Lipid Droplets and Associated Proteins.
abstract: Excess lipid is stored in intracellular organelles known as lipid droplets. This unit discusses techniques for the visualization of lipid droplets and associated proteins in cultured mammalian cells. Protocols for the detection of lipid droplets in fixed or live cells with BODIPY 493/503 are included. The best method for combining visualization of intracellular lipid droplets with indirect immunofluorescent detection of lipid droplet-associated proteins is described. Techniques for sample fixation and permeabilization must be chosen carefully to avoid alterations to lipid droplet morphology. Immunofluorescent detection of perilipin 2, a broadly expressed, lipid droplet-associated protein, widely used as a marker for lipid droplet accumulation, is presented as an example. Finally, a simple protocol for enhancing lipid droplet accumulation through supplementation with excess fatty acid is included. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: Departments of Biology and Chemistry, St. Olaf College, Northfield, Minnesota.
authors.forename: Laura L
authors.initials: LL
authors.lastname: Listenberger
authors.affiliation: Departments of Biology and Chemistry, St. Olaf College, Northfield, Minnesota.
authors.forename: Andrea M
authors.initials: AM
authors.lastname: Studer
authors.affiliation: Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York.
authors.forename: Deborah A
authors.initials: DA
authors.lastname: Brown
authors.affiliation: Center for Human Nutrition, Washington University School of Medicine, St. Louis, Missouri.
authors.forename: Nathan E
authors.initials: NE
authors.lastname: Wolins
mesh_terms: D001896:Boron Compounds; D045744:Cell Line, Tumor; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006801:Humans; D066292:Lipid Droplets / Q000737:chemistry*; D008565:Membrane Proteins / Q000032:analysis*; D000070780:Perilipin-2 / Q000378:metabolism
chemical_list: C527198:4,4-difluoro-1,3,5,7,8-pentamethyl-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D008565:Membrane Proteins; D000070780:Perilipin-2
keywords: BODIPY 493/503; lipid droplet proteins; lipid droplets; perilipin 2