Difference between revisions of "Amplex UltraRed"
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== Signal and output == | == Signal and output == | ||
# Signal: The [[O2k-Fluo LED2-Module]] is operated through the amperometric (Amp)-Channel of the O2k, with electric current (ampere [Amp]) as the primary signal. | # Signal: The [[O2k-Fluo LED2-Module]] is operated through the amperometric (Amp)-Channel of the O2k, with electric current (ampere [Amp]) as the primary signal. | ||
# Output: The focus of the output with Amplex red is on '''[[ | # Output: The focus of the output with Amplex red is on '''[[O2k signal and output| Type B]]''': Flow, flux, rate. | ||
Revision as of 15:14, 28 July 2015
- 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
Signal and output
- Signal: The O2k-Fluo LED2-Module is operated through the amperometric (Amp)-Channel of the O2k, with electric current (ampere [Amp]) as the primary signal.
- Output: The focus of the output with Amplex red is on Type B: Flow, flux, rate.
Different brands
| Trade Mark | Manufacturer/ Distributor | price [€/mg]
|
product id
|
| Amplex Red | 37.4
|
A12222
| |
| Amplex Ultra Red | 48.8
|
A36006
| |
| Ampliflu Red | 18.5
|
90101
| |
| Quanta Red | 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
