Rab7 rfp

Manufactured by Addgene

Rab7-RFP is a fluorescent protein fusion construct that labels the late endosome/lysosome marker Rab7. Rab7 is a small GTPase that regulates trafficking of late endosomes and lysosomes. The Rab7-RFP construct can be used to visualize the localization and dynamics of late endosomes and lysosomes in live cells.

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Lab products found in correlation

Lsm 880 confocal microscope, by Zeiss (2 mentions) Zen 2010, by Zeiss (1 mentions) Dapi fluoromont g, by Southern Biotech (1 mentions) 710 two photon confocal microscope, by Zeiss (1 mentions) 8 well chambered coverglass, by Thermo Fisher Scientific (1 mentions) Lysotracker deep red, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)

2 protocols using rab7 rfp

Visualizing CB2 Receptor Localization

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U2OS
cells transiently transfected with the GFP-tagged CB₂ receptor
(kindly provided by M. Caron and L. S. Barak) and with Rab7-RFP (Addgene,
Cambridge, MA) 48 h earlier were fixed with 4% paraformaldehyde, washed
in phosphate-buffered saline, and mounted with DAPI Fluoromont G (Southern
Biotech, Birmingham, AL). Cells were imaged using a Carl Zeiss 710
two-photon confocal microscope with a 63× oil immersion objective,
using a 1× digital zoom, with excitations set for DAPI, GFP,
and DsRed at 405, 488, and 561 nm, respectively. Images were analyzed
using Zen 2010 (Zeiss), as previously reported.^{16 (link)}

Brailoiu G.C., Deliu E., Marcu J., Hoffman N.E., Console-Bram L., Zhao P., Madesh M., Abood M.E, & Brailoiu E. (2014). Differential Activation of Intracellular versus Plasmalemmal CB2 Cannabinoid Receptors. Biochemistry, 53(30), 4990-4999.

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Lysosome Imaging in PSAP-OE Cells

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Cells were seeded on an 8 well-chambered coverglass (155411, Thermo Fisher Scientific) and imaged on a Carl Zeiss LSM 880 confocal microscope using a 40 × 1.4 NA oil immersion objective. Before the imaging was performed, cell culture media was replaced with phenol red-free media. 50 nM LysoTracker™ Deep Red (L12492, Invitrogen) was directly added to the cell culture media and incubated for 15–60 min before the image acquirement. For RAB5/RAB7 co-localization experiment, RAB5-RFP or RAB7-RFP plasmids (14437 and 14436, Addgene) were transfected to PSAP-OE cells 24 h before the experiment using Lipofectamine 2000 (11668019, Invitrogen) as per manufacturer instructions.

Kojima R., Zurbruegg M., Li T., Paslawski W., Zhang X, & Svenningsson P. (2022). Prosaposin Reduces α-Synuclein in Cells and Saposin C Dislodges it from Glucosylceramide-enriched Lipid Membranes. Journal of Molecular Neuroscience, 72(11), 2313-2325.

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