CTF-LTM 2D Fluorescent Cell Vitality Detection Kit

There are various methods for detecting cell viability, such as detecting dye exclusion, ATP concentration changes, enzyme activity changes, etc. Cell testing sometimes requires studying the mechanisms of cytotoxicity and/or establishing internal reference controls, resulting in the need for multiplex analysis. This involves using multiple analytical methods to detect the same sample, with different detection mechanisms that do not interfere with each other. In this way, relevant comparisons can be made to reveal the mechanisms of cytotoxicity or eliminate systemic differences. Foreign CellTiter Fluor (CTF) products are designed based on this principle. Our company has independently developed and launched a similar fluorescent cell viability assay kit, which has no significant difference in performance compared to foreign brand products. This product is based on the degradation of peptide fluorescent substrate products by proteases in living cells, generating fluorescent signals. When the integrity of the cell membrane is compromised, the protease loses its ability to degrade substrates and does not produce fluorescent signals. CTF-LTMUnlike the luciferase assay kit, which has a different detection principle and method, its detection sensitivity is inferior to the luciferase assay, but it can detect earlier mild cell damage. The two assays have good compatibility and can be used for multiplex detection.
Product Features
1. Performance
Detecting signals, Window， Sensitivity and specificity are similar to similar products from foreign brands.
2、 相容性
Compatible with commonly used analytical reagents such as CTG, without forming mutual influence.
3. Features
More sensitive to early membrane damage in cells.
4. Convenient
The entire experimental procedure of "addition mixed incubation detection" is simple and easy to operate.
Product composition:
Protease substrate, buffer solution mixed and filled into 10 mL or 100 mL brown bottles and 2 mL tubes, with the following specifications:

1. Cell preparation:
(1) Spread cells of appropriate density on 96 well or 384 well cell culture plates for detection. Suggest using a blackboard (if used in combination with luminescent reagents, a whiteboard can be chosen).
(2) Prepare the compound to be tested. Add the appropriate concentration of the compound to be detected to the wells of the cell plate. The concentration of organic solvents in the culture medium should be kept below 1-2%. Establish a control group without added compounds. Continue to cultivate suitable time according to project requirements.
2. Cell viability testing:
(1) Remove CTF-LTMCell viability assay reagent, CTF buffer, was completely dissolved and equilibrated to room temperature. Vortex oscillation of CTF substrate to mix it evenly at the bottom, and instantaneous centrifugation to collect CTF substrate to the bottom of the tube.
(2) Prepare CTF reaction solution: The CTF substrate is 100x, and according to the dosage, prepare 1x reaction solution with CTF buffer and mix thoroughly.
(3) Take out the cell culture plate to be tested and add an equal volume of CTF reaction solution to each well, for example, adding 100 μ l 1x CTF reaction solution to 100 μ l of culture solution.
(4) Mix the low-speed vibrating plate for 20 seconds and react at 37 ℃ in the dark for 60 minutes before conducting fluorescence detection. Different cells have different cutting speeds for CTF substrates, and the optimal plate reading time needs to be optimized according to the cells. The plate reading can be performed as soon as 30 minutes after adding the reagent. Generally, the signal-to-noise ratio of the board reading results is higher after 60 minutes or more. It is recommended to read the board for no more than 3 hours.
(5) Read the fluorescence signal on the fluorescence reader, with excitation light at Ex 380-400nm and emission light at Em 505nm.
(6) Continue downstream multiplex analysis as needed, such as Caspases 3/7 cell apoptosis detection.

1. After initial use, it is recommended to pack the substrate and store it at -20 ℃, and prepare the reaction solution fresh each time.
2. It is not recommended to change the dosage of reaction reagents without strict verification.

-Store in the dark at 20 ℃ and below, with a shelf life of 12 months.