Can I use any of your Cell Meter™ Autophagy Assay Kits with a flow cytometer?

Q: Can I use any of your Cell Meter™ Autophagy Assay Kits with a flow cytometer?

Yes, you can use any of our three Cell Meter™ Autophagy Assay Kits (Cat# 23000, 23001, 23002) with a flow cytometer, fluorescence microscope or fluorescence microplate reader. From our experience Cat# 23002 showed the best response when used in flow cytometric analysis, then Cat# 23001 and finally Cat# 23000. Below is a sample autophagy assay protocol using a flow cytometer: Incubate Jurkat cells in RPMI medium with 10% FBS (control) or in 1X HBSS buffer with 5% FBS (starved) for 16 hours at 37°C. Spin down both the control and starved cells, and then re-suspend in 1X HBSS buffer. Prepare dye working solution by diluting 20 μL of Autophagy Blue™, Autophagy Super Blue™, or Autophagy Green™ (depending on which kit you are using) with 10 mL of Stain Buffer. Stain cells with dye working solution at room temperature for 20-30 minutes. After staining, spin down cells and wash with 1X HBSS buffer. Analyze cells with flow cytometer. Autophagy Blue™ has fluorescence excitation and emission at Ex/Em = 335/520 nm Autophagy Super Blue™ has fluorescence excitation and emission at Ex/Em = 335/520 nm Autophagy Green™ has fluorescence excitation and emission at Ex/Em = 485/530 nm

Question

Can I use any of your Cell Meter&trade; Autophagy Assay Kits with a flow cytometer?

Accepted Answer

Yes, you can use any of our three Cell Meter&trade; Autophagy Assay Kits (Cat# 23000, 23001, 23002) with a flow cytometer, fluorescence microscope or fluorescence microplate reader. From our experience Cat# 23002 showed the best response when used in flow cytometric analysis, then Cat# 23001 and finally Cat# 23000.
Below is a sample autophagy assay protocol using a flow cytometer:

Incubate Jurkat cells in RPMI medium with 10% FBS (control) or in 1X HBSS buffer with 5% FBS (starved) for 16 hours at 37&deg;C.  
Spin down both the control and starved cells, and then re-suspend in 1X HBSS buffer.
Prepare dye working solution by diluting 20 &mu;L of Autophagy Blue&trade;, Autophagy Super Blue&trade;, or Autophagy Green&trade; (depending on which kit you are using) with 10 mL of Stain Buffer.
Stain cells with dye working solution at room temperature for 20-30 minutes.
After staining, spin down cells and wash with 1X HBSS buffer.
Analyze cells with flow cytometer.

Autophagy Blue&trade; has fluorescence excitation and emission at Ex/Em = 335/520 nm
Autophagy Super Blue&trade; has fluorescence excitation and emission at Ex/Em = 335/520 nm
Autophagy Green&trade; has fluorescence excitation and emission at Ex/Em = 485/530 nm

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