|Shipping||Standard overnight for United States, inquire for international|
|H-phrase||H303, H313, H333|
|Intended use||Research Use Only (RUO)|
|R-phrase||R20, R21, R22|
|Storage||Freeze (< -15 °C); Minimize light exposure|
|Excitation||FITC filter set|
|Emission||FITC filter set|
|Recommended plate||Black wall/clear bottom|
|Instrument specification(s)||FITC filter set|
AT A GLANCE
- Prepare and treat cells in a growth medium.
- Incubate cells with Droplite™ Green working solution for 20 to 30 minutes at 37 °C.
- Remove Droplite™ Green working solution.
- Add HHBS buffer and analyze using a fluorescence microscope equipped with a FITC filter set.
The following is our recommended protocol for live cells. This protocol only provides a guideline and should be modified according to your specific needs. Thaw all the kit components at room temperature before starting the experiment.
Plate cells overnight in growth medium at 10,000 to 40,000 cells/well/90 μL for a 96-well plate or 2,500 to 10,000 cells/well/20 μL for a 384-well plate.
Centrifuge the cells from the culture medium.
Suspend the cell pellets in culture medium at 50,000-100,000 cells/well/90 µL for a 96-well poly-D lysine plate or 10,000-25,000 cells/well/20 µL for a 384- well poly-D lysine plate.
Centrifuge the plate at 800 rpm for 2 minutes with the brake off prior to your experiment.
Note: Each cell line should be evaluated on an individual basis to determine the optimal cell density.
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Add 100 µL of DMSO into the Droplite™ Green vial and mix well.
Note: 100 µL stock solution is enough for 100 tests. The staining conditions may be modified according to the particular cell type.
Note: Make a single unused Droplite™ Green stock solution aliquot and store it at ≤ -20 °C. Protect from light and avoid repeated freeze-thaw cycles.
PREPARATION OF WORKING SOLUTION
Add 100 µL of Droplite™ Green stock solution into 10 mL of a buffer of your choice or cell culture medium, and mix well.
Note: HHBS [Hanks' Buffer with 20 mM Hepes] buffer (AAT Cat# 20011) can be used to make a working solution. Prepare a fresh working solution just before use.
SAMPLE EXPERIMENTAL PROTOCOL
Prepare and treat cells in a growth medium as desired.
Add 100 µL/well (96-well plate) of Droplite™ Green working solution to the cell plate.
Note: The optimal concentration of the cell membrane probe varies depending on the specific application.
Incubate the cells at 37 °C for 20 to 30 minutes, protected from light.
Remove the working solution in each well.
Wash twice with DPBS and add HHBS or DPBS solution to the wells.
Observe the fluorescence signal in cells using a fluorescence microscope equipped with a FITC filter set.
Common stock solution preparation
|0.1 mg||0.5 mg||1 mg||5 mg||10 mg|
|1 mM||231.283 µL||1.156 mL||2.313 mL||11.564 mL||23.128 mL|
|5 mM||46.257 µL||231.283 µL||462.567 µL||2.313 mL||4.626 mL|
|10 mM||23.128 µL||115.642 µL||231.283 µL||1.156 mL||2.313 mL|
|Mass (Calculate)||Molecular weight||Volume (Calculate)||Concentration (Calculate)||Moles|
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