Talk:Iglesias-Gonzalez 2013 J Neurosci Methods

Ice-cold isolation buffer used in study

225 mM Mannitol

75 mM Sucrose

5 mM HEPES,

1 mg/ml fatty acid free BSA (pH 7.4, isotonized with KOH)

• EGTA or EDTA added at concentrations in the range of: 1 - 3 mM

• Final mitochondrial pellet was resusbended in IB without chelators.

• Respiratory assays were carried out as previously described in Iglesias-Gonzalez 2012 Neurochem Res:

Standard respiration medium contains:

125 mM KCl

5 mM HEPES

3 mM MgCl₂

2 mM KH₂PO₄

0,5 mM EGTA (pH 7.4)

Chemicals for MiR05/MiR06

Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.

Compound	Final conc.	FW	Addition to 1 litre final volume	Company, product code and storage
EGTA	0.5 mM	380.4	0.190 g	Sigma E 4378, 25 g, store at R.T.
MgCl2.6H20	3 mM	203.3	0.610 g	Scharlau MA 0036, 250 g, store at R.T.
Taurine	20 mM	125.1	2.502 g	Sigma T 0625, 25 g, store at R.T.
KH2PO4	10 mM	136.1	1.361 g	Merck 104873, 1000 g, store at R.T.
HEPES	20 mM	238.3	4.77 g	Sigma H 7523, 250 g, store at R.T.
D-Sucrose	110 mM	342.3	37.65 g	Roth, 4621.1, 1000 g, store at R.T.
BSA essentially fatty acid free	1 g/l		1 g	Sigma A 6003 fraction V, 25 g, store at 2-8 °C
Lactobionic acid	60 mM	358.3	120 ml of 0.5 M K-lactobionate stock	Aldrich 153516 or Sigma L2398, 100 g, store at R.T.

MiRK03

>> from intern wiki MiRK03

KCl ? mM, adjust osmolarity (official: osmotic concentration) to MiR05

HEPES free acid 20 mM

KH₂PO₄ 10 mM

MgCl₂ 3 mM

EGTA 0.5 mM

BSA: 0.1%

pH = 7.0

KCl 130 mM