Amplex UltraRed

high-resolution terminology - matching measurements at high-resolution

Amplex UltraRed

Description

Amplex red (AmR) is used as an extrinsic fluorophore for measurement of hydrogen peroxide production (ROS) by cells or mitochondrial preparations. The reaction of H₂O₂ and AmR is catalyzed by horseradish peroxidase to produce the red fluorescent compound resorufin (excitation wavelength 563 nm, emission 587 nm). The change of emitted fluorescence intensity is directly proportional to the concentration of H₂O₂ added, whereby the H₂O₂ is consumed.

Abbreviation: AmR

Reference: Mohanty 1997 J Immunol Methods, Zhou 1997 Anal Biochem, Mishin 2010 Free Radical Biol Med, Towne 2004 Anal Biochem, Krumschnabel 2015 Methods Mol Biol

MitoPedia methods: Fluorometry 

Different brands

Trade Mark	Manufacturer/ Distributor	price [€/mg]	product id
Amplex Red	Life Technologies	37.4	A12222
Amplex Ultra Red	Life Technologies	48.8	A36006
Ampliflu Red	Sigma	18.5	90101
Quanta Red	Thermo scientific		15150

Citation	Amplex		HRP			pH	Limit of detection
	stock	final	unit definition	stock	final
	mM	µM		U/ml	U/ml
BIOTEK	10	50	pyrogallol	10	0.1	7.4	4 nM (absorption 300 nm)
Life Technologies	10	50		10	0.1	6 to 7.5	<80 nM
Towne 2004		160			0.41	7.5 to 8.5	100 nM
Zhou1997		3 ?			0.3 to 1		50 nM (10 nM optimal)
Mohanty 1997	100	10 to 100			1		18 nM
Komary 2010		1			2.5

Applications with O2k-Fluorometry

Source: Life Technologies (former Invitrogen) A36006

• Krumschnabel G, Fontana-Ayoub M, Sumbalova Z, Heidler J, Gauper K, Fasching M, Gnaiger E (2015) Simultaneous high-resolution measurement of mitochondrial respiration and hydrogen peroxide production. Methods Mol Biol 1264:245-61. »Bioblast pdf«

O2k-Demo experiments

• MiPNet20.14 AmplexRed H2O2-production
• MiPNet18.05_Amplex-Mouse-heart
• MiPNet18.06_Amplex-Yeast