Image-iT™ Green Hypoxia Reagent, 1 Vial - FAQs

View additional product information for Image-iT™ Green Hypoxia Reagent - FAQs (I14833, I14834)

7 product FAQs found

Can Image-it Green Hypoxia Reagent be used in cells expressing fluorescent proteins?

Yes, as long as the fluorescent protein does not emit in the green range (˜510–535 nm). For cells that do not express a fluorescent protein, we recommend viewing unstained cells under the FITC channel 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 the FITC channel.

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

Our Image-iT Green Hypoxia Reagent is already highly fluorescent before using the product. What could have caused this?

This could be due to storage of the product under anaerobic or low oxygen conditions. The Image-iT Green Hypoxia Reagent increases in fluorescence upon exposure to low oxygen environments, and this change is not reversible. For some ROS indicators, we recommend storing the reagent under dry nitrogen or argon to prevent oxidation during storage. However, this is not appropriate for the Image-iT Green Hypoxia Reagent.

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

What is different between the use of the Hypoxia Green Reagent for Flow Cytometry (Cat No. H20035) versus imaging using the Image-iT Green Hypoxia Reagent (Cat Nos. I14833, I14834)?

The difference between these products is in the final working concentration of the reagent and incubation time. For flow cytometry applications, we recommend using a final concentration in the range of 0.5 to 1 µM with an incubation time ranging from 2 to 3 hrs. For imaging, we recommend a final concentration in the range of 1 to 10 µM with an incubation time ranging from 30 mins to 1 hr. For both applications, one should optimize the final working concentration and incubation time.

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

Is Hypoxia Green Reagent for Flow Cytometry (Cat No. H20035) the same reagent as the Image-iT Green Hypoxia Reagent (Cat Nos. I14833, I14834)?

The Hypoxia Green Reagent for Flow Cytometry and the Image-iT Green Hypoxia Reagent are the same reagent, formulated differently for flow cytometry and imaging use, respectively.

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

May I include a total cell stain (to stain all cells) along with Image-iT Hypoxia Reagent?

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.

May I include a dead cell stain along with Image-iT Hypoxia Reagent?

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.

Which filter set can I use for optimal excitation and emission with Image-iT Green Hypoxia Reagent?

The Image-iT Green Hypoxia Reagent has an approximate Ex/Em maxima at 488/520 nm. We recommend using a 488 nm laser and a FITC/GFP emission filter.

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