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| == Application in [[HRR]] ==
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| '''MiR06: Mitochondrial Respiration Medium - MiR06''' (MiR06 = MiR05 + Catalase); Total volume = 1 litre.
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| <span style="color:#8B008B"> '''Caution:''' Chemicals stored in the fridge or freezer should be adjusted to room temperature before opening.</span>
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| '''Preparation of 500 mM ADP stock solution''' (dissolved in H<sub>2</sub>O):
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| ::1) weigh 491.2 mg of ADP (1 mmol ADP)
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| ::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!
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| ::3) neutralize with 5 N KOH (approx. 465 µl) !ADP will dissolve after addition of KOH! (this is '''Solution A''')
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| ::4) check pH and adjust to 7 if necessary
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| ::5) adjust final volume to 2 ml and divide into 0.2 ml portions
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| ::6) store frozen at -80 °C
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| '''Preparation of 500 mM ADP stock solution with 3 mM free Mg<sup>2+</sup>''':
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| 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).
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| '''MgCl<sub>2</sub>''' (Scharlau MA0036: MgCl<sub>2</sub>.6H<sub>2</sub>O, FW = 203.3)
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| ::1) add 121,98 mg MgCl<sub>2</sub> to '''Solution A'''
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| ::2) follow steps 4-6 of the above instructions
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| '''Oxygraph-2k Manual Titrations:''' [[MiPNet09.12]]
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| ::* Titration volume: 8-20 µl using a 25 µl syringe
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| ::* Final conc. in 2 ml O2k-chamber: 2-5 mM
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Revision as of 17:56, 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.
