Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Blue Fluorescence Optimized for Flow Cytometry*

Additional fluorescent color(s): 
Image Viewer
Detection of hydrogen peroxide in Jurkat cells using Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit (Cat#: 11505). Jurkat cells were stained with OxiVision™ Blue peroxide sensor for 30 minutes and treated with 100 µM hydrogen peroxide at 37 °C for 90 minutes. Cells stained with OxiVision™ Blue peroxide sensor but without hydrogen peroxide treatment were used as control.
Roll over image to zoom in
Loading...
 
Unit Size: Cat No: Price (USD): Qty:
100 Tests 11505 $245


Export item/cart as Excel file

Send item/cart as email
EXPORT TO EXCEL X

Export:
EXPORT TO EMAIL X
Important: We request your email address to ensure that the recipient(s) knows you intended for them to see the email, and that it is not junk mail.
Export:
Your Name*:
Your Email*:
Recipient Email*:
Your Personal Message:
Additional Ordering Information
Telephone: 1-800-990-8053
Fax: 1-408-733-1304
Email: sales@aatbio.com
International: See distributors





Overview

Ex/Em (nm)405/450
Storage Freeze (<-15 °C)
Minimize light exposure
InstrumentsFlow cytometer
Category Enzyme Detection
Horseradish Peroxidase (HRP)
Related Reactive Oxygen Species
Microplate Readers
Hydrogen peroxide is a reactive oxygen metabolic by-product that serves as a key regulator for a number of oxidative stress-related states. It is involved in many biological events that are linked to asthma, atherosclerosis, diabetic vasculopathy, osteoporosis, a number of neurodegenerative diseases and Down's syndrome. The measurement of this reactive species is helpful for determining how oxidative stress modulates various intracellular pathways. This Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit uses our unique OxiVision™ Blue peroxide sensor to quantify hydrogen peroxide in live cells. OxiVision™ Blue peroxide sensor is cell-permeable, and generates blue fluorescence when it reacts with hydrogen peroxide. This kit provides a sensitive tool to monitor hydrogen peroxide level in living cells, and it is optimized to be used in flow cytometry.




Protocol


Quick Preview

This protocol only provides a guideline, and should be modified according to your specific needs.
At a glance

Protocol summary

  1. Prepare cells in growth medium
  2. Stain cells with OxiVision™ Blue Peroxide Sensor
  3. Treat cells with test compounds
  4. Monitor fluorescence intensity with flow cytometer Pacific Blue Channel (Ex/Em = 405/450 nm)

Important notes
Thaw kit components at room temperature before starting the experiment.

Key parameters
Instrument:Flow cytometer
Excitation:Pacific Blue channel
Emission:Pacific Blue channel
Preparation of stock solution
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

1. OxiVision™ Blue Peroxide Sensor stock solution:
Add 100 µL of DMSO (Component B) into the vial of OxiVision™ Blue peroxide sensor (Component A), and mix them well. Note: 1 µL of reconstituted OxiVision™ Blue peroxide sensor stock solution is enough for 0.5 mL cells. The stock solution should be used promptly. Keep from light.

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

Sample experimental protocol
  1. Stain cells with OxiVision™ Blue Peroxide Sensor stock solution in full medium or in your desired buffer at 37°C for 20 - 30 minutes, protected from light.

  2. Treat cells with test compounds in full medium or in your desired buffer at 37°C for desired period of time. For control samples (untreated cells), add the corresponding amount of compound buffer. Note: It’s recommended to treat cells in full medium. However, if tested compounds are serum sensitive, growth medium and serum factors can be aspirated away before treatment. Resuspend cells in 1X Hank’s salt solution and 20 mM Hepes buffer (HHBS) or the buffer of your choice after aspiration. Alternatively, cells can be treated in serum-free media. Note: We treated Jurkat cells with 100 µM hydrogen peroxide in full medium at 37°C for 90 minutes to induce hydrogen peroxide. See Figure 1 for details.

  3. Monitor the fluorescence intensity at Pacific Blue channel (Ex/Em=405/450 nm) using a flow cytometer. Gate on the cells of interest, excluding debris.
Example data analysis and figures

Figure 1. Detection of hydrogen peroxide in Jurkat cells using Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit (Cat#: 11505). Jurkat cells were stained with OxiVision™ Blue peroxide sensor for 30 minutes and treated with 100 µM hydrogen peroxide at 37 °C for 90 minutes. Cells stained with OxiVision™ Blue peroxide sensor but without hydrogen peroxide treatment were used as control.
Disclaimer
AAT Bioquest provides high-quality reagents and materials for research use only. For proper handling of potentially hazardous chemicals, please consult the Safety Data Sheet (SDS) provided for the product. Chemical analysis and/or reverse engineering of any kit or its components is strictly prohibited without written permission from AAT Bioquest. Please call 408-733-1055 or email info@aatbio.com if you have any questions.





References & Citations

Plasma-activated medium selectively eliminates undifferentiated human induced pluripotent stem cells
Authors: Ryo Matsumoto, Kazunori Shimizu, Takunori Nagashima, Hiromasa Tanaka, Masaaki Mizuno, Fumitaka Kikkawa, Masaru Hori, Hiroyuki Honda
Journal: Regenerative Therapy (2016): 55--63






Additional Documents

 
Safety Data Sheet (SDS)


Catalogs
1. Reactive Oxygen Species (ROS) Detection

Certificate of Analysis