Talk:Iglesias-Gonzalez 2013 J Neurosci Methods: Difference between revisions
From Bioblast
No edit summary |
No edit summary |
||
Line 1: | Line 1: | ||
'''Ice-cold isolation buffer used in study''' | |||
225 mM Mannitol | 225 mM Mannitol |
Revision as of 15:06, 28 January 2013
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