CD63 is a membrane marker tetraspanin protein of EVs, and we prepared HeLa cells stably expressing GFP-fused CD63 to secrete CD63-GFP-containing EVs (CD63-GFP-EVs) as previously reported9 (link)10 (link). HeLa cells (1 × 105 cells) were plated on a 24-well microplate (Iwaki, Tokyo, Japan) and incubated for 1 day. They were transfected with CD63-GFP plasmid (pCT-CD63-GFP, pCMV, Cyto-Tracer, System Biosciences, Mountain View, CA) (800 ng) complexed with Lipofectamine LTX reagent (2 μl) and PLUS reagent (1 μl) (Invitrogen, Life Technologies Corporation) in α-MEM containing 10% FBS (200 μl). The cells were also treated with puromycin (3 μg/ml) (LKT Laboratories, St. Paul, MN) for the antibiotic selection of HeLa cells stably expressing CD63-GFP (CD63-GFP-HeLa).
Cyto tracer
Manufactured by System Biosciences
Sourced in United States, Canada
The Cyto-Tracer is a specialized lab equipment designed for cellular analysis. It is used to track and monitor the movement and behavior of cells in a controlled environment.
Automatically generated - may contain errors
3 protocols using cyto tracer
1
Generating Stable CD63-GFP-Expressing HeLa Cells
2
Generation of CD63-GFP-Exosomes in HeLa Cells
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Tetraspanin CD63 is a membrane marker protein of exosomes. We prepared HeLa cells stably expressing GFP-fused CD63 to secrete CD63-GFP-containing exosomes (CD63-GFP-exosomes). HeLa cells (4.7 × 104 cells) were plated on a 24-well microplate (Iwaki, Tokyo, Japan) and incubated for 1 day. The cells were transfected with CD63-GFP plasmid (pCT-CD63-GFP, pCMV, Cyto-Tracer, System Biosciences, Mountain View, CA, USA; 800 ng) complexed with Lipofectamine LTX reagent (2 μl) with PLUS reagent (1 μl; Invitrogen, Life Technologies Corporation) in α-MEM containing 10% FBS (200 μl). The cells were also treated with puromycin (3 μg/ml; LKT Laboratories, St. Paul, MN, USA) for the antibiotic selection of HeLa cells stably expressing CD63-GFP (CD63-GFP-HeLa).
3
Generating Exosomes with CD63-GFP Fusion
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Tetraspanin CD63 is a membrane marker protein of exosomes, and we prepared HeLa cells stably expressing GFP-fused CD63 to secrete CD63-GFP-containing exosomes (CD63-GFP-exosomes). The cells (1 × 105 cells) were plated on a 24 well microplate (Iwaki, Tokyo, Japan) and incubated for 1 day. They were transfected with CD63-GFP plasmid (pCT-CD63-GFP, pCMV, Cyto-Tracer, System Biosciences, Mountain View, CA) (800 ng) complexed with Lipofectamine LTX reagent (2 μl) with PLUS reagent (1 μl) (Invitrogen, Life Technologies Corporation) in α-MEM containing 10% FBS (200 μl). The cells were also treated with puromycin (3 μg/ml) (LKT Laboratories, St. Paul, MN) for the antibiotic selection of HeLa cells stably expressing CD63-GFP (CD63-GFP-HeLa).
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