Thermo Scientific™ Hypoxia Green Reagent for Flow Cytometry

Catalog No.	Quantity
H20035	2 x 50 assays

Catalog No. H20035 Supplier Thermo Scientific™ Supplier No. H20035

Description

Hypoxia Green Reagent for Flow Cytometry is an antibody-independent reagent used to detect low levels of oxygen levels in live cells.

The membrane-permeant probe releases rhodamine as oxygen levels decrease, resulting in a fluorogenic response. Hypoxia is a characteristic of many diseases, including cardiovascular disease and tumor-mediated immunosuppression and is critical to tumor survival and growth.

• Sensitive, fixed, end-point assay
• Detects decreases in oxygen in live cells
• Fluorogenic, non-antibody based probe

Specifications

• Content And Storage: 2 vials of 50 tests each
  Store at -5°C to -30°C.
• Detection Method: Fluorescence
• For Use With (Equipment): Flow Cytometer
• Product Type: Reagent
• Emission: 488
• Format: Vial(s)
• Quantity: 2 x 50 assays
• Shipping Condition: Room Temperature

Frequently Asked Questions (FAQs)

Can the Hypoxia Green Reagent for Flow Cytometry 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.

Our Hypoxia Green Reagent for Flow Cytometry 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 Hypoxia Green Reagent for Flow Cytometry 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 Hypoxia Green Reagent for Flow Cytometry.

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.

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.