6-FAM SE [92557-81-8]

Katalog-Nummer HY-15939-10mg

Size : 10mg

Marke : MedChemExpress

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Beschreibung

6-FAM SE (6-carboxyfluorescein succinimidyl ester) is a fluorescent labeling reagent. 6-FAM SE is used for oligonucleotide labeling and DNA sequencing[1].

In Vitro

1. Sample preparation[1]:
1) Collect cells or biological samples to be stained. For cell samples, ensure that the cells are in the logarithmic growth phase. After collecting the cells, wash them twice with an appropriate buffer (such as PBS) to remove serum and other impurities in the culture medium.
2) Count the cells using a cell counter or hemocytometer and adjust the cell concentration to the 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 the required concentration. Generally speaking, the molecular weight of 6-FAM SE is about 467.4.
2) Dissolve 6-FAM SE powder in anhydrous DMSO to prepare a high concentration stock solution, usually 1-5 mM. For example, to prepare a 1 mM stock solution, dissolve 467.4 μg of 6-FAM SE in 1 mL of DMSO. Dispense the stock solution into sterile microcentrifuge tubes to avoid repeated freezing and thawing. It can be stored at -20℃ or lower.
3. Preparation of working solution:
Before use, take out a tube of dye stock solution and thaw it at room temperature. Dilute the stock solution to the required working concentration, generally 0.5-5 μM, with cell culture medium or buffer preheated to 37℃. For example, take 10 μL of 1 mM stock solution and add it to 2 mL of culture medium to obtain 5 μM working solution.
4. Specific staining steps:
1) Mix the prepared cell sample with the dye working solution in an appropriate ratio. Generally, the volume ratio of cell suspension to working solution is 1:1 or 1:2. For example, take 1 mL of cell suspension and add 1-2 mL of dye working solution.
2) Mix gently to allow the cells to fully contact the dye and incubate in an incubator at 37°C and 5% CO2 for 15-30 min. Gently shake or invert several times during this period to ensure uniform staining.
3) After incubation, wash the 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 and use them for subsequent experimental analysis, such as flow cytometry detection, fluorescence microscopy observation, etc.

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

Lösungsmittel & Löslichkeit
In Vitro: 

DMSO : 50 mg/mL (105.62 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.1124 mL 10.5621 mL 21.1242 mL
5 mM 0.4225 mL 2.1124 mL 4.2248 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.5 mg/mL (5.28 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.5 mg/mL (5.28 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Reinheit & Dokumentation
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HY-15939-50mg
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HY-15939-100mg
 100mg