- 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
