Amplex™ Red Glutamic Acid/Glutamate Oxidase Assay Kit
Amplex™ Red Glutamic Acid/Glutamate Oxidase Assay Kit
Invitrogen™

Amplex™ Red Glutamic Acid/Glutamate Oxidase Assay Kit

The Amplex™ Red Glutamic Acid/Glutamate Oxidase Assay Kit provides a sensitive and simple method for detecting glutamic acid/glutamate oxidase activityRead more
Have Questions?
Catalog number A12221
Price (USD)
458.00
Each
Add to cart
Price (USD)
458.00
Each
Add to cart
The Amplex™ Red Glutamic Acid/Glutamate Oxidase Assay Kit provides a sensitive and simple method for detecting glutamic acid/glutamate oxidase activity using a fluorescence microplate reader or fluorometer.

See our complete line of Fluorescence Microplate assays.

• Detect concentrations as low as 10 nM L-glutamic acid or 40 μU/mL of purified L-glutamate oxidase
• Format allows for multiple time point measurements
• Designed for minimal autofluorescence interference

The Amplex™ Red Glutamic Acid/Glutamate Oxidase Assay Kit provides an ultrasensitive method for detecting glutamic acid or for continuously monitoring glutamate oxidase activity using a fluorescence microplate reader or fluorometer. In the assay, L-glutamic acid is oxidized by glutamate oxidase to produce α-ketoglutarate, NH3, and hydrogen peroxide. L-alanine and L-glutamate–pyruvate transaminase are included in the reaction to regenerate L-glutamic acid by transamination of α-ketoglutarate, resulting in multiple cycles of the initial reaction and a significant amplification of the hydrogen peroxide produced. The hydrogen peroxide reacts with 10-acetyl-3,7-dihydroxyphenoxazine (Amplex™ Red reagent) in a 1:1 stoichiometric ratio in a reaction catalyzed by horseradish peroxidase (HRP) to generate the highly fluorescent product resorufin.

Because resorufin has absorption and fluorescence emission maxima of approximately 571 nm and 585 nm, respectively, there is little interference from autofluorescence in most biological samples.

Use Amplex™ Red Assays for a Broad Range of Investigations
A wide variety of validated Amplex™ Red assays are available for studying cell signaling and lipids, neurobiology, inflammation and immune function, and metabolism. We also offer Amplex™ UltraRed Reagent (Cat. No. A36006), a second-generation reagent providing greater sensitivity and brighter fluorescence, and the Amplex™ Red/UltraRed Stop Reagent (Cat. No. A33855). The Amplex™ Red/UltraRed Stop Reagent provides convenience and control by allowing the fluorescence signal-generating reaction to be terminated at a user-determined time point. After addition of the stop reagent, the fluorescence signal remains stable for at least three hours. Custom assay design and packaging are also available.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodFluorescence
For Use With (Application)Glutamic Acid/Glutamate Oxidase Assay
For Use With (Equipment)Fluorometer, Microplate Reader
Product TypeRed Glutamic Acid/Glutamate Oxidase Assay Kit
Dye TypeOther Label(s) or Dye(s)
Format96-well plate
Product LineAmplex™
Quantity200 assays
Shipping ConditionRoom Temperature
Unit SizeEach
Contents & Storage
Store in freezer -5°C to -30°C and protect from light.

Frequently asked questions (FAQs)

I'm using an Amplex Red kit, the reagent changes color to pink almost immediately in my own Krebs-Ringer buffer but not in HBSS. Why is this?

The components of Krebs-Ringer buffer (salts) should not cause oxidation of the Amplex reagent (which, in the presence of peroxidase and H2O2 oxidizes to resorufin, which is pink in color and fluorescent). Try water alone (the water used to make the Krebs-Ringer buffer). Since Hank's Buffered Saline Solution is typically purchased rather than made in the lab, it likely would not have the same contaminant. Another option is to degas the buffer prior to use to removed dissolved oxygen radicals.

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

Can Amplex Red Assays be performed using cell lysates?

This is not recommended. The presence of endogenous proteases can complicate the assay by degrading the horseradish peroxidase (HRP). Endogenous peroxidases and antioxidants can modify the H2O2 required for the reaction, competing with HRP (and catalase) for the substrate.

The Amplex Red Assays are best performed with either purified enzymes or extracted H2O2 in a defined buffer system, extracellular solutions or body fluids (media, serum, etc.) that do not exhibit high levels of endogenous protease or oxidase activity and do not contain antioxidants.

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