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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Lab products found in correlation

Nsg1 emerald, by Addgene (1 mentions) Mcherry rab5ca q79l, by Addgene (1 mentions) Mcherry rab5dn s34n, by Addgene (1 mentions) Orca flash 4, by Hamamatsu Photonics (1 mentions) Inverted spinning disk confocal microscope, by PerkinElmer (1 mentions) Plan apo vc, by Nikon (1 mentions)

2 protocols using egfp rab7

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.

Yap C.C., Digilio L., McMahon L, & Winckler B. (2017). The endosomal neuronal proteins Nsg1/NEEP21 and Nsg2/P19 are itinerant, not resident proteins of dendritic endosomes. Scientific Reports, 7, 10481.

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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.

Kounatidis I., Stanifer M.L., Phillips M.A., Paul-Gilloteaux P., Heiligenstein X., Wang H., Okolo C.A., Fish T.M., Spink M.C., Stuart D.I., Davis I., Boulant S., Grimes J.M., Dobbie I.M, & Harkiolaki M. (2020). 3D Correlative Cryo-Structured Illumination Fluorescence and Soft X-ray Microscopy Elucidates Reovirus Intracellular Release Pathway. Cell, 182(2), 515-530.e17.

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