Invitrogen™ Image-iT™ Red Hypoxia Reagent

Catalog No.	Quantity
H10498	1 mg

Catalog No. H10498 Supplier Invitrogen™ Supplier No. H10498

Description

Image-iT™ Red Hypoxia Reagent is a novel fluorogenic compound for measuring hypoxia in live cells.

It is non-fluorescent when live cells are in an environment with normal oxygen concentrations and becomes fluorescent when oxygen levels are decreased. Image-iT™ Red Hypoxia Reagent is a real-time oxygen detector, with a fluoregenic response that reverses when the cells return to normal oxygen levels.

Features of Image-iT™ Red Hypoxia Reagent include:

Measures hypoxia in live cells by fluorescing in low oxygen environments

Real-time oxygen detector, with reversible fluoregenic response

Easy to use—just add to cell culture media and image

Image-iT™ Red Hypoxia Reagent is a fluorogenic compound that becomes fluorescent in environments with low oxygen concentrations, and it is live cell permeable. These properties makes it a highly useful tool for detecting cells and tissue under hypoxic conditions. Image-iT™ Red Hypoxia Reagent is a very sensitive oxygen detector. Unlike pimonidazole adducts that respond only to very low oxygen levels, Image-iT™ Hypoxia Reagent begins to fluoresce when atmospheric oxygen levels are less than 5%. It responds quickly to such environments, and the fluorogenic response reverses if oxygen concentrations improve. These properties make Image-iT™ Red Hypoxia Reagent an ideal tool for detecting hypoxic conditions around tumors, 3D cultures, spheroids, neurons, etc. It can be used to detect tumors in small animals, and its fluorogenic properties have been shown to corresponds with increased Hif 1α expression and translocation in hypoxic environments (1).

1. Zhang, S., Hosaka, M., Yoshihara, T., Negishi, K., Iida, Y., Tobita, S., & Takeuchi, T. (2010) Phosphorescent light–emitting iridium complexes serve as a hypoxia-sensing probe for tumor imaging in living animals. Cancer Research 70(11), 4490-4498.

Specifications

• Content And Storage: Provided as 1 mg lyophilized solid. Enough for 3000 assays with 0.1 mL/assay or 600 assays with 0.5 mL/assay
  When stored at ≤-20°C product is stable for (at least 6 months after receipt.
  Protect from light and desiccate.)
• Detection Method: Fluorescence
• Product Type: Fluorogenic Reagent
• Form: Solid
• Product Line: Image-iT
• Quantity: 1 mg
• Reagent Type: Oxygen Sensor
• Shipping Condition: Approved for shipment at Room Temperature or on Wet Ice

Frequently Asked Questions (FAQs)

Can Image-iT Red Hypoxia Reagent be used in cells expressing fluorescent proteins?

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.

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.

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.

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

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).