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AUTOdot Autophagy Visualization Dye

Monodansylpentane Cadaverine Staining Tool

Localization of guayule Small Rubber Particle Protein (SRPP-GFP) to lipid droplets (LD) in a tobacco cell. Shown are representative epifluorescence micrographs of tobacco (Nicotiana tabacum) Bright Yellow-2 (BY-2) cells, which serve as a well-characterized system for studying protein localization in plant cells. BY-2 cells have been transiently transformed via biolistic bombardment with plasmid DNA-encoding full-length guayule SRPP12 C-terminally fused to the N-terminus of the Green Fluorescent Protein (SRPP-GFP). Following bombardment, cells have been incubated in linoleic acid, which induces an increase in the number and size of LD in these cells, and then incubated with AUTODOTTM, which is a blue-fluorescing marker dye for LD in living cells. The fluorescence attributable to the AUTODOTTM stained LD is false colorized red. The yellow color in the merged images represents obvious co-localizations between SRPP-GFP and AUTODOTTM -stained LD, most of which have coalesced, due to the ectopic overexpression of the fusion protein. These larger coalesced structures are not observed in the neighboring non-transformed cells wherein LD are usually dispersed throughout the cytosol. Similar coalescence of LD has been observed in BY-2 cells transiently overexpressing Arabidopsis LDAP, as well as in various other cells types in which other LD proteins, such as Perilipin-1 and the Ancient Ubiquitous Protein-1, are ectopically overexpressed. Shown also is the corresponding differential interference contrast image. Bar = 10 μm. Plant Signaling & Behavior 8, e27141; 2013
Image of untreated and autophagy induced mouse cerebral cells analyzed by fluorescence microscopy using an inverted microscope equipped with a filter system (excitation filer: 380-420 nm, barrier filter: 450 nm).

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Description	AUTODOT™ preferentially segregates into the neutral lipid cores of LDs and emits blue fluorescence, compatible with concurrent use of green and red fluorescent reporters in live-cell imaging. It can be used for visualizing LDs in cell cultures and fixed tissues, making it a versatile marker for LDs in fluorescence microscopy. Major lipid-based pathways such as autophagy, lipolysis, fatty acid oxidation, ketogenesis, and cholesterol synthesis are amenable to tracking by AUTODOT™.
Concentration	0.1M
Target/Specificity	AUTOdot is a monodansylpentane (MDH) staining tool specific for autopahgic vacuoles.
Format	Product is 0.1M MDH supplied in DMSO.
Storage	Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	AUTOdot Autophagy Visualization Dye is for research use only and not for use in diagnostic or therapeutic procedures.