6-CFDA is a common aliphatic luciferin-line organism. CFDA conducts free diffusion into cells, and then it is hydrolyzed into carboxyl fluorescein (CF) by intracellular non-specific lipase.
Preparation of 6-CFDA working solution
1.Preparation of the stock solution
Dissolve 1mg 6-CFDA in 0.2172 mL DMSO to obtain 10 mM of 6-CFDA.
Note: It is recommended to store the stock solution at -20 °C -80 °C away from light and avoid repetitive freeze-thaw cycles.
2.Preparation of 6-CFDA working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of 6-CFDA working solution.
Note: Please adjust the concentration of 6-CFDA working solution according to the actual situation.
Cell staining
1.Cell preparation:
For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.Add 1 mL of 6-CFDA working solution, and then incubate at room temperature for 30 minutes.
3.Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
4.Wash twice with PBS, 5 minutes each time.
5.Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.
Precautions
1. It is recommended to store the stock solution at -20℃ or -80℃ away from light and avoid repetitive freeze-thaw cycles.
2. Please adjust the concentration of 6-CFDA working solution according to the actual situation.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.
| Molecular Weight | 460.39 |
| Formula | C25H16O9 |
| CAS Number | 3348-03-6 |
| Solubility (25°C) | DMSO 90 mg/mL |
| Storage | -20°C, protect from light |
| Species | Mouse | Rat | Rabbit | Guinea pig | Hamster | Dog |
| Weight (kg) | 0.02 | 0.15 | 1.8 | 0.4 | 0.08 | 10 |
| Body Surface Area (m2) | 0.007 | 0.025 | 0.15 | 0.05 | 0.02 | 0.5 |
| Km factor | 3 | 6 | 12 | 8 | 5 | 20 |
| Animal A (mg/kg) = Animal B (mg/kg) multiplied by | Animal B Km |
| Animal A Km |
For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.
| Related Fluorescent Dye Products |
|---|
| SOSG
SOSG (Singlet Oxygen Sensor Green Fluorescent Probe) is highly selective for singlet oxygen (1O2). It is commonly used for the detection of singlet oxygen in solutions and the evaluation of the efficacy of free radical scavengers. |
| FM1-43
FM1-43 is a very lipophilic, water-soluble styrene dyes, can specifically bind to cell membranes and inner membrane organelles to produce fluorescence. FM1-43 is widely used in endocytic and exospic membrane structure markers. |
| BCIP
BCIP is a chromogenic substrate for colorimetric assay of alkaline phosphatase activity. |
| MCA-SEVNLDAEFR-K(Dnp)-RR, amide
MCA-SEVNLDAEFR-K(Dnp)-RR, amide is a FRET-based substrate. |
| MeOSuc-AAPV-AMC
MeOSuc-AAPV-AMC is a fluorogenic substrate for human leukocyte and porcine pancreatic elastase (Km: 362 μM, Ex=380 nm, Em=460 nm). |
Products are for research use only. Not for human use. We do not sell to patients.
© Copyright 2010-2023 AbMole BioScience. All Rights Reserved.
