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Egfp rab7

Manufactured by Addgene

EGFP-Rab7 is a fluorescent protein fusion construct that consists of the enhanced green fluorescent protein (EGFP) fused to the Rab7 protein. Rab7 is a small GTPase that regulates late endosome and lysosome trafficking. The EGFP tag allows for the visualization of the localization and dynamics of Rab7 within the cell.

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2 protocols using egfp rab7

1

Fluorescent Protein-Tagged Rab GTPases

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Nsg1-Emerald:Addgene cat#54202, Michael Davidson lab; the mutation at residue 114 from D to G was corrected.
Nsg1-mCherry:mouse full length Nsg1 was cloned at the N-terminal of mCherry in pcDNA3 vector.
Nsg2-GFP:mouse full length Nsg2 was cloned into pcDNA-GFP by Genscript
mRFP-Rab5:Addgene cat#14437, Ari Helenius lab.
mCherry Rab5CA(Q79L) cat#35138, mCherry Rab5DN(S34N) cat#35139:Addgene, Sergio Grinstein lab.
EGFP-Rab7 cat#12605, EGFP-Rab7DN(T22N) cat#12660:Addgene, Richard Pagano labmTagBFP2-Rab5, cat#56417, Addgene, Michael Davidson lab;
mTagBFP2-Rab7:Michael Davidson lab.
GFP:Clontech
mCherry:Clontech.
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2

Live-cell Imaging of Rab7 and MRV

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U2OS wild-type or stably expressing mCherry-Gal3 were transfected with an eGFP-Rab7 (Addgene) expressing plasmid 16 h prior to imaging. MRV labeled with Alexa647 was added to cells and then imaging was started. Live-cell imaging was performed with an inverted spinning-disk confocal microscope (PerkinElmer) using oil immersion objectives (60x, 1.49 NA, Apo TIRF, Nikon or 100x, 1.4 NA, Plan Apo VC, Nikon) and a CMOS camera (Orca Flash 4, Hamamatsu). Cells, objectives and microscope stage were kept at 37°C and 5% CO2 through the presence of an environment-control chamber. Cells were imaged in 0.5 μM stacks 5min apart for 180 min.
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