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Description
Mitochondrial respiration medium, MiRK03, modified after a medium described by Komary 2010 Biochim Biophys Acta, intended for use as medium for H2O2 production measurement with Amplex Red.
Abbreviation: MiRK03
Reference: Krumschnabel 2014 Methods Mol Biol, Komary 2010 Biochim Biophys Acta
MitoPedia topics: Media for respirometry
MiRK03
substance | MiRK03 | molecular weight | for 1 l |
---|---|---|---|
EGTA | 0.5 mM | 380.35 g/mol | 190.18 mg |
MgCl2 x 6 H2O | 3 mM | 203.30 g/mol | 609.90 mg |
HEPES | 20 mM | 238.31 g/mol | 4 766.2 mg |
KCl | 130 mM | 74.56 g/mol | 9 692.8 mg |
KH2PO4 | 10 mM | 136.09 g/mol | 1 360.9 mg |
BSA | 1 mg/ml | - | 1000 mg |
Preparation without BSA
- Weigh all ingredients (besides BSA) according to the table above and transfer them into a big glass beaker.
- Add about 750 mL H2O.
- Under stirring adjust the temperature to 25 ยฐC.
- Adjust the pH to 7.12 with 5 M KOH.
- Adjust final volume to 1 L and divide in 40 mL portions.
- Store at -20 ยฐC.
MiRK03 ph - temperature dependence
data recorded with MiRK03 without BSA (only averaged values are shown; two measurements were conducted)
temperature [ยฐC] | pH of MiRK03 |
---|---|
20 | 7.15 |
25 | 7.12 |
30 | 7.09 |
35 | 7.05 |
37 | 7.03 |
- For comparison see: Mitochondrial respiration media: comparison