6-FAM SE

Guide (The following is our recommended experimental protocol. This protocol is for reference only and should be modified according to your specific needs).
1. Sample preparation^[1]:
1. Collect cells or biological samples to be stained. For cell samples, ensure cells are in the logarithmic growth phase. After collection, wash cells twice with an appropriate buffer (such as PBS) to remove serum and other impurities from the culture medium.
2. Count cells using a cell counter or hemocytometer, and adjust the cell concentration to an appropriate range.
2. Preparation of dye stock solution:
1. Weigh an appropriate amount of 6-FAM SE powder and calculate the required mass based on its molecular weight and desired concentration. Generally, the molecular weight of 6-FAM SE is approximately 473.39.
2. Dissolve 6-FAM SE powder in anhydrous DMSO to prepare a high-concentration stock solution, typically 1-5 mM. For example, to prepare a 1 mM stock solution, dissolve 473.39 μg of 6-FAM SE in 1 mL of DMSO. Aliquot the stock solution into sterile microcentrifuge tubes, avoid repeated freeze-thaw cycles, and store at -20°C or lower.
3. Preparation of working solution:
Before use, take out one tube of dye stock solution and thaw at room temperature. Dilute the stock solution with pre-warmed (37°C) cell culture medium or buffer to the desired working concentration, typically 0.5-5 μM. For example, add 10 μL of a 1 mM stock solution to 2 mL of medium to obtain a 5 μM working solution.
4. Staining procedure:
1. Mix the prepared cell sample with the dye working solution at an appropriate ratio, typically a volume ratio of cell suspension to working solution of 1:1 or 1:2. For example, add 1-2 mL of dye working solution to 1 mL of cell suspension.
2. Gently mix to ensure full contact between cells and dye, and incubate at 37°C in a 5% CO₂ incubator for 15-30 min. During incubation, gently shake or invert several times to ensure uniform staining.
3. After incubation, wash cells at least twice with pre-cooled buffer to remove unbound dye. After each wash, centrifuge at an appropriate speed (such as 300-500 g) for 5-10 min and discard the supernatant.
4. Finally, resuspend the stained cells in an appropriate amount of buffer or culture medium for subsequent experimental analysis, such as flow cytometry or fluorescence microscopy.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.