Difference between revisions of "MiR06"

Latest revision as of 16:23, 8 February 2016

high-resolution terminology - matching measurements at high-resolution

MiR06

Description

Mitochondrial respiration medium, MiR06, developed for oxygraph incubations of mitochondrial preparations. MiR06 = MiR05 plus catalase. MiR06Cr = MiR06 plus creatine.

Abbreviation: MiR06

Reference: MiPNet14.13 Medium-MiR06

MitoPedia topics: Media for respirometry

@@ Line 1: / Line 1: @@
 {{MitoPedia
 |abbr=MiR06
-|description='''Mitochondrial respiration medium 06''', developed for oxygraph incubations of [[mitochondrial preparations]] = MiR05 plus catalase.
+|description='''Mitochondrial respiration medium, [[MiPNet14.13 Medium-MiR06|MiR06]]''', developed for oxygraph incubations of [[mitochondrial preparations]].  MiR06 = MiR05 plus [[catalase]]. MiR06Cr = MiR06 plus [[creatine]].
-|info=[[MiPNet14.13]]; [[Gnaiger 2000 MitoInTheCold]]
+|info=[[MiPNet14.13 Medium-MiR06]]
 |type=Chemicals
 }}
-{{Labeling
+{{MitoPedia concepts|type=Chemicals
-|instruments=Chemicals; Media
+}}
+{{MitoPedia methods|type=Chemicals
+}}
+{{MitoPedia O2k and high-resolution respirometry|type=Chemicals
+}}
+{{MitoPedia topics
+|mitopedia topic=Medium
 |type=Chemicals
 }}
-== Application in [[HRR]] ==
-'''MiR06: Mitochondrial Respiration Medium - MiR06''' (MiR06 = MiR05 + Catalase); Total volume = 1 litre.
-<span style="color:#8B008B"> '''Caution:''' Chemicals stored in the fridge or freezer should be adjusted to room temperature before opening.</span>
-'''Preparation of 500 mM ADP stock solution''' (dissolved in H<sub>2</sub>O):
-::1) weigh 491.2 mg of ADP (1 mmol ADP)
-::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!
-::3) neutralize with 5 N KOH (approx. 465 µl)  !ADP will dissolve after addition of KOH!  (this is '''Solution A''')
-::4) check pH and adjust to 7 if necessary
-::5) adjust final volume to 2 ml and divide into 0.2 ml portions
-::6) store frozen at -80 °C
-'''Preparation of 500 mM ADP stock solution with 3 mM free Mg<sup>2+</sup>''':
-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).
-'''MgCl<sub>2</sub>''' (Scharlau MA0036: MgCl<sub>2</sub>.6H<sub>2</sub>O, FW = 203.3)
-::1) add 121,98 mg MgCl<sub>2</sub> to '''Solution A'''
-::2) follow steps 4-6 of the above instructions
-'''Oxygraph-2k Manual Titrations:'''  [[MiPNet09.12]]
-::* Titration volume: 8-20 µl using a 25 µl syringe
-::* Final conc. in 2 ml O2k-chamber: 2-5 mM