Thank you for reading my questions!
I want to do the permeability assay for the Caco-2 cells on the transwell-like structure.
I have several questions:
(1) What is the general concentration for the test?
(2) How to make the stocking solution and how is the store conditions? The FITC-dextran is easily dissolved into PBS, could I dissolve them, and prepare 1mg/ml stocking solution, store at -25℃.
(3) For calibration under spectrometer, what are general concentrations?
I hope someone who has done the permeability assay, could give me some suggestions.
Thank you very much!
Based on my experience, I only answer questions (1) and (2).
(1) Permeability was measured using a 1 mg/mL FITC-dextran solution.
(2) Stocking solution was dissolved in HBSS at a concentration of 100 mg/mL and stored at 4 ℃. PBS can also be used regardless.
The transferred FITC-dextran concentration was determined with excitation and emission wavelengths of 490 and 520 nm.
For question 3, the calibration of the spectrometer depends on the purpose of the experiment and which spectrometer you are using. I used a dilution of the FITC solution based on 1mg/mL.
Hi!
Currently I'm working with caco2 monolayer cells and I have performed FITC Permeability at different timepoints.
I would like to know how have you interpreted the data? I used a standard conc ranging from 100 ug/ml to 1 ug/ml and having HBBS solution as blank. The absorbance values goes till 1000 which is in line with the standard conc.
Could you please let me know the procedure? If you have a raw data for the same, that would be useful.
Thanks!
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line