1 mg FITC Dextran (Cat#T18988, Targetmol, USA) was dissolved in 1 ml double-distilled water to obtain a stock solution with a concentration of 1 mg/ml. The stock solution was diluted with serum-free medium to a working concentration of 500 μg/ml. After this intervention, PBS was added for washing twice. 100 μl FITC Dextran (500 μg/ml) was added to the upper compartment, and 600 μl serum-free basal medium was added to the lower compartment. After 1 h incubation in the dark, 100 μl medium was aspirated from each compartment and loaded onto a black 96-well plate. The OD value was measured at 495nm/520nm using a multi-functional enzyme reader (MD, USA). The permeability was calculated [25 (link)].
Fitc dextran
Manufactured by Targetmol
Sourced in United States
FITC Dextran is a fluorescently labeled polysaccharide commonly used in various biomedical and biochemical applications. It serves as a tracer and marker for studying molecular transport, permeability, and diffusion processes in biological systems.
Automatically generated - may contain errors
2 protocols using fitc dextran
1
FITC Dextran Permeability Assay
2
Cytotoxicity and Cellular Uptake of NAP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
WST-1 assay (Beyotime) was used to determine the cytotoxicity of NAP. In brief, A549 cells (2 × 103) were seeded in 96-well plate and treated with increasing concentration of NAP (up to 400 μM) for 48 h. WST-1 was added and the plate was read after 1.5 h on an automated microplate spectrophotometer (BioTek Instruments, Vermont, USA) at 450 nm according to the manufacturer’s instructions.
Colony-formation assay was performed to detect long-term survival of A549 cells in NAP treatment. 400 cells seeded in 12-well plates were treated with indicated concentration of NAP for 15 days. The plates were washed twice with PBS and fixed with methanol for 10 min at room temperature and then stained with 1% crystal violet for 5 min. The number of colonies was counted using ImageJ software (version 1.44I).
Cellular uptake assay was performed by using FITC-Dextran (TargetMol). A549 cells were seeded onto 12-well plate and incubated with FITC-Dextran (0.1 mg/mL) for 24 h, then the cells were treated with 50, 100, 150 and 200 μM of NAP for 24 h and 48 h, respectively. The fluorescence (excited at 488 nm and emitted at 530 nm) was measured with a multifunctional microplate reader Spark 10 M (Tecan, Synergy-HT).
Colony-formation assay was performed to detect long-term survival of A549 cells in NAP treatment. 400 cells seeded in 12-well plates were treated with indicated concentration of NAP for 15 days. The plates were washed twice with PBS and fixed with methanol for 10 min at room temperature and then stained with 1% crystal violet for 5 min. The number of colonies was counted using ImageJ software (version 1.44I).
Cellular uptake assay was performed by using FITC-Dextran (TargetMol). A549 cells were seeded onto 12-well plate and incubated with FITC-Dextran (0.1 mg/mL) for 24 h, then the cells were treated with 50, 100, 150 and 200 μM of NAP for 24 h and 48 h, respectively. The fluorescence (excited at 488 nm and emitted at 530 nm) was measured with a multifunctional microplate reader Spark 10 M (Tecan, Synergy-HT).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!