Difference between revisions of "MiR06"
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== Application in [[HRR]] == | |||
'''MiR06: Mitochondrial Respiration Medium - MiR06''' (MiR06 = MiR05 + Catalase); Total volume = 1 litre. | |||
<span style="color:#8B008B"> '''Caution:''' Chemicals stored in the fridge or freezer should be adjusted to room temperature before opening.</span> | |||
'''Preparation of 500 mM ADP stock solution''' (dissolved in H<sub>2</sub>O): | |||
::1) weigh 491.2 mg of ADP (1 mmol ADP) | |||
::2) add 1.3 ml H<sub>2</sub>O !White precipitate will occur, which will be dissolved after 1-2 min of stirring on magnetic stirrer at room temperature! | |||
::3) neutralize with 5 N KOH (approx. 465 µl) !ADP will dissolve after addition of KOH! (this is '''Solution A''') | |||
::4) check pH and adjust to 7 if necessary | |||
::5) adjust final volume to 2 ml and divide into 0.2 ml portions | |||
::6) store frozen at -80 °C | |||
'''Preparation of 500 mM ADP stock solution with 3 mM free Mg<sup>2+</sup>''': | |||
To keep free [MgMg<sup>2+</sup>] constant during respiration measurement mix ADP (or ATP) with MgCl<sub>2</sub> (0.6 mol/mol ADP or 0.8 mol/mol ATP). | |||
'''MgCl<sub>2</sub>''' (Scharlau MA0036: MgCl<sub>2</sub>.6H<sub>2</sub>O, FW = 203.3) | |||
::1) add 121,98 mg MgCl<sub>2</sub> to '''Solution A''' | |||
::2) follow steps 4-6 of the above instructions | |||
'''Oxygraph-2k Manual Titrations:''' [[MiPNet09.12]] | |||
::* Titration volume: 8-20 µl using a 25 µl syringe | |||
::* Final conc. in 2 ml O2k-chamber: 2-5 mM | |||
Revision as of 14:53, 22 November 2010
- high-resolution terminology - matching measurements at high-resolution
MiR06
Description
Mitochondrial respiration medium 06, developed for oxygraph incubations of mitochondrial preparations = MiR05 plus catalase.
Abbreviation: MiR06
Reference: MiPNet14.13; Gnaiger 2000 MitoInTheCold
Labels:
HRR: Chemicals; Media"Chemicals; Media" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property.
Application in HRR
MiR06: Mitochondrial Respiration Medium - MiR06 (MiR06 = MiR05 + Catalase); Total volume = 1 litre.
Caution: Chemicals stored in the fridge or freezer should be adjusted to room temperature before opening.
Preparation of 500 mM ADP stock solution (dissolved in H2O):
- 1) weigh 491.2 mg of ADP (1 mmol ADP)
- 2) add 1.3 ml H2O !White precipitate will occur, which will be dissolved after 1-2 min of stirring on magnetic stirrer at room temperature!
- 3) neutralize with 5 N KOH (approx. 465 µl) !ADP will dissolve after addition of KOH! (this is Solution A)
- 4) check pH and adjust to 7 if necessary
- 5) adjust final volume to 2 ml and divide into 0.2 ml portions
- 6) store frozen at -80 °C
Preparation of 500 mM ADP stock solution with 3 mM free Mg2+:
To keep free [MgMg2+] constant during respiration measurement mix ADP (or ATP) with MgCl2 (0.6 mol/mol ADP or 0.8 mol/mol ATP).
MgCl2 (Scharlau MA0036: MgCl2.6H2O, FW = 203.3)
- 1) add 121,98 mg MgCl2 to Solution A
- 2) follow steps 4-6 of the above instructions
Oxygraph-2k Manual Titrations: MiPNet09.12
- Titration volume: 8-20 µl using a 25 µl syringe
- Final conc. in 2 ml O2k-chamber: 2-5 mM
