Fluorescent Protein Transfection Protocol

FusionRed-F-tractin has been previously described [3 (link)]. EGFP-F-tractin (no. 58473), mEmerald (no. 53975), LifeAct-mEmerald (no. 54148), mScarlet-I (no. 85044), and LifeAct-mScarlet-I (no. 85054) were obtained from Addgene (Watertown, MA). 1 μg of plasmid was used to transfect 750,000 cells in each well of a 6-well plate using Lipofectamine 2000 (5 μL; Thermo Fisher Scientific) in OptiMEM (400 μL; Thermo Fisher Scientific). After 30 min at room temperature, plasmid in Lipofectamine 2000/OptiMEM was then incubated with cells in complete media (2 mL) overnight.

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Vosatka K.W., Lavenus S.B, & Logue J.S. (2022). A novel Fiji/ImageJ plugin for the rapid analysis of blebbing cells. PLoS ONE, 17(4), e0267740.

Publication 2022

mEmerald LifeAct-mEmerald mScarlet-I Lipofectamine 2000 OptiMEM

Cells Lipofectamine 2000 Plasmid

Corresponding Organization : Albany Medical Center Hospital

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Variable analysis

independent variables

Plasmid used for transfection: FusionRed-F-tractin, EGFP-F-tractin, mEmerald, LifeAct-mEmerald, mScarlet-I, LifeAct-mScarlet-I

dependent variables

Not explicitly mentioned

control variables

Number of cells transfected: 750,000 cells per well of a 6-well plate
Transfection reagent: Lipofectamine 2000 (5 μL)
Transfection medium: OptiMEM (400 μL)
Incubation time: 30 min at room temperature
Complete media: 2 mL

controls

No positive or negative controls were explicitly mentioned.

Annotations

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