MiRK03: Difference between revisions

Latest revision as of 10:01, 9 November 2020

high-resolution terminology - matching measurements at high-resolution

MiRK03

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 °C.
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 °C.

MiRK03 ph - temperature dependence

data recorded with MiRK03 without BSA (only averaged values are shown; two measurements were conducted)

temperature [°C]	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

@@ Line 1: / Line 1: @@
 {{MitoPedia
 |abbr=MiRK03
-|description='''Mitochondrial respiration medium, MiRL03''', modified after a medium described by [[Komary 2010 Biochim Biophys Acta]], intended for use as medium for H2O2 production measurement with Amplex Red.
+|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.
 |info=[http://bioblast.at/index.php/Krumschnabel_2014_Methods_Mol_Biol Krumschnabel 2014 Methods Mol Biol], [http://bioblast.at/index.php/Komary_2010_Biochim_Biophys_Acta Komary 2010 Biochim Biophys Acta]
+|type=Chemicals
+}}
+{{MitoPedia concepts|type=Chemicals
+}}
+{{MitoPedia methods|type=Chemicals
 }}
-{{MitoPedia methods
+{{MitoPedia O2k and high-resolution respirometry|type=Chemicals
-|mitopedia method=Respirometry, Fluorometry
 }}
 {{MitoPedia topics
@@ Line 11: / Line 15: @@
 |type=Chemicals
 }}
-* For composition see: [[Mitochondrial respiration medium]]
+=== MiRK03 ===
+{| class="wikitable"
+|-
+! substance !! MiRK03 !! molecular weight !! for 1 l
+|-
+| EGTA || 0.5 mM|| 380.35 g/mol || 190.18 mg
+|-
+| MgCl<sub>2</sub> x 6 H<sub>2</sub>O|| 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
+|-
+| KH<sub>2</sub>PO<sub>4</sub>|| 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 H<sub>2</sub>O.
+::::# Under stirring adjust the temperature to 25 °C.
+::::# 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 °C.
+==== MiRK03 ph - temperature dependence ====
+data recorded with ''MiRK03 without BSA'' (only averaged values are shown; two measurements were conducted)
+{| class="wikitable"
+|-
+! temperature [°C] !! 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]]