Image-iT™ Red Hypoxia Reagent - FAQs

Yes, as long as the fluorescent protein does not emit in the red range (˜610 nm). For cells that do not express a fluorescent protein, we recommend viewing unstained cells under the TRITC and Texas Red channels to examine autofluorescence. For cells expressing a fluorescent protein, we recommend analyzing unstained cell samples to determine the extent the fluorescent protein emission may overlap in these channels.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

You may use cell-permeable nuclear counterstains or surface labels, but avoid any general cytoplasmic stains that may interfere with the emission of the hypoxia reagents.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Yes. We recommend using NucGreen Dead 488 ReadyProbes Reagent (Cat No. R37109) with Image-iT Red Hypoxia Reagent and NucRed Dead 647 ReadyProbes Reagent (Cat No. R37113) with Image-iT Green Hypoxia Reagent.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

The Image-iT Red Hypoxia Reagent has an approximate Ex/Em maxima at 490/610 nm. You may use a 488 nm laser and a Texas Red emission filter. For lamp-based instruments, a YFP longpass filter set may be used (excitation range from 485 to 510 nm and emission from 550 nm up to 620 nm).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.