Difference between revisions of "Catalase"
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:::: '''Unit definition:''' '''* Units''' of enzymatic activity (U) in µmol/min; assay used by Sigma Aldrich: ' ''One unit will decompose 1.0 μmole of H<sub>2</sub>O<sub>2</sub> per min at pH 7.0 at 25 °C, while the H<sub>2</sub>O<sub>2</sub> concentration falls from 10.3 to 9.2 mM, measured by the rate of decrease of A<sub>240</sub>.'' ' | :::: '''Unit definition:''' '''* Units''' of enzymatic activity (U) in µmol/min; assay used by Sigma Aldrich: ' ''One unit will decompose 1.0 μmole of H<sub>2</sub>O<sub>2</sub> per min at pH 7.0 at 25 °C, while the H<sub>2</sub>O<sub>2</sub> concentration falls from 10.3 to 9.2 mM, measured by the rate of decrease of A<sub>240</sub>.'' ' | ||
:::: Use 'units/mg solid' for your calculations. | |||
:::: Example: 'Catalase lypophilized powder; 2,800 U/mg solid and 3,500 units/mg protein'. | |||
:::: '''Preparation of stock solution: 112,000 U/mL ''' (dissolved in MiR05) for '''2-mL O2k-chamber''': | :::: '''Preparation of stock solution: 112,000 U/mL ''' (dissolved in MiR05) for '''2-mL O2k-chamber''': | ||
::::# Result: 40 mg catalase powder (2,800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 112,000 U/mL. | ::::# Result: 40 mg catalase powder (2,800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 112,000 U/mL. | ||
::::# Divide into 0.2 mL portions. | ::::# Divide into 0.2 mL portions. | ||
| Line 30: | Line 32: | ||
::::* Titration volume ('''2-mL O2k-chamber'''): 5 µL using a 25 µL Hamilton syringe. | ::::* Titration volume ('''2-mL O2k-chamber'''): 5 µL using a 25 µL Hamilton syringe. | ||
::::* Final concentration: 280 U/mL. | |||
:::: '''Preparation of stock solution: 28,000 U/mL ''' (dissolved in MiR05) for '''0.5-mL O2k-chamber''': | |||
::::# Use 'units/mg solid' for your calculations. Example: 'Catalase lypophilized powder; 2,800 U/mg solid and 3,500 units/mg protein'. | |||
::::# 10 mg catalase powder (2,800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 28,000 U/mL. | |||
::::# Divide into 0.2 mL portions. | |||
::::# Store frozen at -20 °C. | |||
:::»'''O2k manual titrations:''' [[MiPNet09.12]] | |||
::::* Titration volume ('''0.5-mL O2k-chamber'''): 5 µL using a 10 µL Hamilton syringe. | |||
::::* Final concentration: 280 U/mL. | ::::* Final concentration: 280 U/mL. | ||
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=== Preparation of catalase stock solution === | === Preparation of catalase stock solution === | ||
::'''Stock solution:''' 112,000 U/mL (dissolved in MiR05) | ::'''Stock solution:''' 112,000 U/mL (dissolved in MiR05) | ||
Revision as of 10:34, 9 November 2020
- high-resolution terminology - matching measurements at high-resolution
Catalase
Description
Catalase catalyzes the dismutation of hydrogen peroxide to water and oxygen. Perhaps all cells have catalase, but mitochondria of most cells lack catalase. Cardiac mitochondria are exceptional in having mt-catalase activity (rat heart mitochondria: Radi et al 1991; mouse heart mitochondria: Rindler et al 2013).
Abbreviation: Ctl
Reference: Radi 1991 J Biol Chem, Rindler 2013 J Biol Chem, MiPNet09.12
MitoPedia topics: Enzyme
Communicated by Gnaiger E (2015-03-05).
Application in HRR
- Clt: Catalase (from bovine liver, lyophilized powder; 2,000-5,000 U/mg protein); Sigma C 9322; store at -20 °C.
- Catalase is included in the mitochondrial respiration medium MiR06 (MiR06 = MiR05 plus catalase), so as to allow for re-oxygenation of the respiration medium (without lifting the stopper and risking the introduction of air bubbles) by addition of a small volume of a 200 mM stock solution of H2O2.
- Unit definition: * Units of enzymatic activity (U) in µmol/min; assay used by Sigma Aldrich: ' One unit will decompose 1.0 μmole of H2O2 per min at pH 7.0 at 25 °C, while the H2O2 concentration falls from 10.3 to 9.2 mM, measured by the rate of decrease of A240. '
- Use 'units/mg solid' for your calculations.
- Example: 'Catalase lypophilized powder; 2,800 U/mg solid and 3,500 units/mg protein'.
- Preparation of stock solution: 112,000 U/mL (dissolved in MiR05) for 2-mL O2k-chamber:
- Result: 40 mg catalase powder (2,800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 112,000 U/mL.
- Divide into 0.2 mL portions.
- Store frozen at -20 °C.
- »O2k manual titrations: MiPNet09.12
- Titration volume (2-mL O2k-chamber): 5 µL using a 25 µL Hamilton syringe.
- Final concentration: 280 U/mL.
- Preparation of stock solution: 28,000 U/mL (dissolved in MiR05) for 0.5-mL O2k-chamber:
- Use 'units/mg solid' for your calculations. Example: 'Catalase lypophilized powder; 2,800 U/mg solid and 3,500 units/mg protein'.
- 10 mg catalase powder (2,800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 28,000 U/mL.
- Divide into 0.2 mL portions.
- Store frozen at -20 °C.
- Preparation of stock solution: 28,000 U/mL (dissolved in MiR05) for 0.5-mL O2k-chamber:
- »O2k manual titrations: MiPNet09.12
- Titration volume (0.5-mL O2k-chamber): 5 µL using a 10 µL Hamilton syringe.
- Final concentration: 280 U/mL.
