Difference between revisions of "Amplex UltraRed"
From Bioblast
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{{MitoPedia | {{MitoPedia | ||
|abbr=AmR | |abbr=AmR | ||
|description='''Amplex red''' (AmR) | |description='''Amplex red''' (AmR) is used as an [[extrinsic fluorophores |extrinsic fluorophore]] for measurement of [[hydrogen peroxide]] production ([[ROS]]) by cells or mitochondrial preparations. The reaction of H<sub>2</sub>O<sub>2</sub> 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 H<sub>2</sub>O<sub>2</sub> added, whereby the H<sub>2</sub>O<sub>2</sub> is consumed. | ||
|info=[[Mohanty 1997 J Immunol Methods]], [[Zhou 1997 Anal Biochem]], [[Mishin 2010 Free Radical Biol Med]], [[Towne 2004 Anal Biochem]], [[Krumschnabel 2015 Methods Mol Biol]] | |info=[[Mohanty 1997 J Immunol Methods]], [[Zhou 1997 Anal Biochem]], [[Mishin 2010 Free Radical Biol Med]], [[Towne 2004 Anal Biochem]], [[Krumschnabel 2015 Methods Mol Biol]] | ||
}} | }} | ||
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{{MitoPedia topics}} | {{MitoPedia topics}} | ||
== Different brands == | |||
{| style="border-spacing:0;" | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| '''Trade Mark''' | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| '''Manufacturer/ Distributor''' | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <div align="right">'''<nowiki>price [€/mg]</nowiki>'''</div> | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <div align="right">'''product id'''</div> | |||
|- | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| Amplex Red | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <center>Life Technologies</center> | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <div align="right">37.4</div> | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <div align="right">A12222</div> | |||
|- | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| Amplex Ultra Red | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <center>Life Technologies</center> | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <div align="right">48.8</div> | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <div align="right">A36006</div> | |||
|- | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| Ampliflu Red | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <center>Sigma</center> | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <div align="right">18.5</div> | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <div align="right">90101</div> | |||
|- | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| Quanta Red | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <center>Thermo scientific</center> | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| | |||
| style="border:none;padding-top:0cm;padding-bottom:0cm;padding-left:0.053cm;padding-right:0.053cm;"| <div align="right">15150</div> | |||
|} | |||
{| border="1" style="border-spacing:0;" style="text-align: center;" | |||
|| Citation | |||
| colspan="2" | Amplex | |||
| colspan="3" | HRP | |||
|| pH | |||
|| Limit of detection | |||
|- | |||
|| | |||
|| stock | |||
|| final | |||
|| unit definition | |||
|| stock | |||
|| final | |||
|| | |||
|| | |||
|- | |||
|| | |||
|| <div align="right">mM</div> | |||
|| <div align="right">µM</div> | |||
|| | |||
|| <div align="right">U/ml</div> | |||
|| <div align="right">U/ml</div> | |||
|| | |||
|| | |||
|- | |||
|| BIOTEK | |||
|| <div align="right">10</div> | |||
|| <div align="right">50</div> | |||
|| <div align="right">pyrogallol</div> | |||
|| <div align="right">10</div> | |||
|| <div align="right">0.1</div> | |||
|| <div align="right">7.4</div> | |||
|| <div align="right">4 nM (absorption 300 nm)</div> | |||
|- | |||
|| Life Technologies | |||
|| <div align="right">10</div> | |||
|| <div align="right">50</div> | |||
|| | |||
|| <div align="right">10</div> | |||
|| <div align="right">0.1</div> | |||
|| <div align="right">6 to 7.5</div> | |||
|| <div align="right"><nowiki><80 nM</nowiki></div> | |||
|- | |||
|| [[Towne_2004_Anal_Biochem|Towne 2004]] | |||
|| | |||
|| <div align="right">160</div> | |||
|| | |||
|| | |||
|| <div align="right">0.41</div> | |||
|| <div align="right">7.5 to 8.5</div> | |||
|| <div align="right">100 nM</div> | |||
|- | |||
|| [[Zhou_1997_Anal_Biochem|Zhou1997]] | |||
|| | |||
|| <div align="right">3 ?</div> | |||
|| | |||
|| | |||
|| <div align="right">0.3 to 1</div> | |||
|| | |||
|| <div align="right">50 nM (10 nM optimal)</div> | |||
|- | |||
|| [[Mohanty_1997_J_Immunol_Methods |Mohanty 1997]] | |||
|| <div align="right">100</div> | |||
|| <div align="right">10 to 100</div> | |||
|| | |||
|| | |||
|| <div align="right">1</div> | |||
|| | |||
|| <div align="right">18 nM</div> | |||
|- | |||
|| [[Komary 2010 Biochim Biophys Acta |Komary 2010]] | |||
|| | |||
|| <div align="right">1</div> | |||
|| | |||
|| | |||
|| <div align="right">2.5</div> | |||
|| | |||
|| | |||
|} | |||
== Applications with O2k-Fluorometry == | == 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. [[Krumschnabel 2015 Methods Mol Biol |»Bioblast pdf«]] | ||
:* [[MiPNet18.05_Amplex-Mouse-heart]] | |||
:* [[ | '''O2k-Demo experiments''' | ||
::* [[MiPNet19.20 AmplexRed H2O2-production]] | |||
::* [[MiPNet18.05_Amplex-Mouse-heart]] | |||
::* [[MiPNet18.06_Amplex-Yeast]] |
Revision as of 15:19, 7 June 2015
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 | 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