Cells were grown overnight in 24-well plates at 37°C in a 5% CO2 atmosphere. After reaching over 80% confluence, the plasmids mRuby-Clathrin (Addgene #55852), pEGFP-Sec23A (Addgene #66609), ZsGreen-Rab5 (custom synthesized by Genomeditech (Shanghai, China) Co., Ltd.), EGFP-Rab7A (Addgene #28047), GFP-LAMP1 (Addgene #16290), EMTB-3XGFP (Addgene #26741), pSNAPf-Cox8A (Addgene #101129), or pSNAPf-TOMM20 (custom synthesized by Genomeditech (Shanghai, China) Co., Ltd.) was transfected into the cells using Lipofectamine 3000 (Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. After 24 h, the transfected cells were digested with trypsin-EDTA and seeded into Nunc Glass Bottom Dishes (Φ 12 mm, Thermo Fisher Scientific, Inc.) at a density of 1.5 ~ 2.0 × 104 per well in growth medium (150 μL). The cells were grown for an additional 12 ~ 24 h before incubation with the indicated probes.
Zsgreen rab5
Manufactured by Addgene
ZsGreen-Rab5 is a fluorescently labeled protein construct that consists of the ZsGreen fluorescent protein fused to the Rab5 protein. Rab5 is a small GTPase that regulates early endosome formation and trafficking. The ZsGreen-Rab5 fusion protein can be used to visualize and track the movement of early endosomes within live cells.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 3000, by Thermo Fisher Scientific (4 mentions)
Lamp1 gfp, by Addgene (2 mentions)
Mruby clathrin, by Addgene (2 mentions)
Pegfp sec23a, by Addgene (2 mentions)
Psnapf tomm20, by Thermo Fisher Scientific (2 mentions)
Egfp rab7a, by Addgene (2 mentions)
Psnapf cox8a, by Addgene (2 mentions)
Nunc glass bottom dishes, by Thermo Fisher Scientific (2 mentions)
Emtb 3xgfp, by Addgene (2 mentions)
2 protocols using zsgreen rab5
1
Fluorescent Protein Transfection Workflow
2
Fluorescent protein-tagged cell imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were grown overnight in 24-well plates at 37°C in a 5% CO 2 atmosphere. After reaching over 80% confluence, the plasmids mRuby-Clathrin (Addgene #55852), pEGFP-Sec23A (Addgene #66609), ZsGreen-Rab5 (custom synthesized by Genomeditech (Shanghai, China) Co., Ltd.), EGFP-Rab7A (Addgene #28047), GFP-LAMP1 (Addgene #16290), EMTB-3XGFP (Addgene #26741), pSNAPf-Cox8A (Addgene #101129), or pSNAPf-TOMM20 (custom synthesized by Genomeditech (Shanghai, China) Co., Ltd.) was transfected into the cells using Lipofectamine 3000 (Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. After 24 h, the transfected cells were digested with trypsin-EDTA and seeded into Nunc Glass Bottom Dishes (Φ 12 mm, Thermo Fisher Scientific, Inc.) at a density of 1.5~2.0 × 104 per well in growth medium (150 µL). The cells were grown for an additional 12~24 h before incubation with the indicated probes.
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