ICT’s Basic Calcein AM Cell Viability Kit allows researchers to easily and simultaneously differentiate between live and dead cells within a single sample. To use Calcein AM, simply add the reagent directly to the cell sample, incubate, and analyze (no wash steps necessary). Samples can be analyzed using a flow cytometer, fluorescence plate reader, or fluorescent microscope.
- Prepare samples and controls.
- Dilute 10X Cellular Assay Buffer 1:10 with diH2O.
- Reconstitute Calcein AM with 50 µL DMSO to prepare 2 mM stock solution.
- Dilute 2 mM Calcein AM stock solution 1:5 by adding 200 µL diH2O or PBS, forming a 400 µM stock solution.
- Stain with Calcein AM at a concentration between 1-10 µM. The ideal staining concentration can vary based on cell line, application, etc., and should be determined by the end user. The recommended sample size is 0.4 mL.
- Add Calcein AM to each sample and mix gently. For example, to stain a 0.4 mL sized sample with 10 µM Calcein AM, add 10 µL of the 400 µM stock solution.
- Incubate approximately 1 hour.
- Analzye with a flow cytometer, fluorescence microscope, or fluorescence plate reader. Calcein AM excites at 494 nm and emits at 520 nm.