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Difference between revisions of "Cytochrome c"

From Bioblast
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== Application in [[HRR]] ==
== Application in [[HRR]] ==


''' ''c'': Cytochrome ''c'' ''' (L-Glutamic acid, sodium salt, C<sub>5</sub>H<sub>8</sub>NO<sub>4</sub>Na; contains 1 mol/mol H<sub>2</sub>O); Sigma G 1626 (R.T.): FW = 187.1 or 169.1 anhydrous)
''' ''c'': Cytochrome ''c'' ''' (Sigma C 7752, 50mg, store at -20 °C): FW = 12500)
 
'''Preparation of 4 mM Cytochrome ''c'' solution''' (dissolved in H<sub>2</sub>O):


'''Preparation of 2 M L-Glutamic acid solution''' (dissolved in H<sub>2</sub>O):
Chemicals stored in the fridge or the freezer should be adjusted to room temperature beforing opening!


::1) weigh 1.871 g of L-Glutamic acid and transfer to 5 mL volumetric glass flask
::1) weigh 50 mg of Cytochrome ''c'' into a small glass vial. Difficult to weigh, since the powder appears to be electrostatic.
::2) add 3 ml H<sub>2</sub>O (pH = 6)
::2) add 1 ml H<sub>2</sub>O. Dissolves easily.
::3) neutralize with 5 N KOH (approx. 50 µl)
::3) divide into 0.2 ml portions  
::4) check pH and adjust to 7 if necessary 
::4) store frozen at -20 °C
::5) adjust final volume to 5 ml and divide into 0.5 ml portions.
::6) store frozen at -20 °C


'''Oxygraph-2k Manual Titrations:'''  [[MiPNet09.12]]
'''Oxygraph-2k Manual Titrations:'''  [[MiPNet09.12]]


::* Titration volume: 10 µl using a 25 µl syringe
::* Titration volume: 5 µl using a 25 µl syringe
::* Final conc. in 2 ml O2k-chamber: 10 mM
::* Final conc. in 2 ml O2k-chamber: 10 µM

Revision as of 10:34, 22 November 2010


high-resolution terminology - matching measurements at high-resolution


Cytochrome c

Description

Abbreviation: c

Reference: MiPNet09.12; Gnaiger_2002_BiochemSocTrans


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Application in HRR

c: Cytochrome c (Sigma C 7752, 50mg, store at -20 °C): FW = 12500)

Preparation of 4 mM Cytochrome c solution (dissolved in H2O):

Chemicals stored in the fridge or the freezer should be adjusted to room temperature beforing opening!

1) weigh 50 mg of Cytochrome c into a small glass vial. Difficult to weigh, since the powder appears to be electrostatic.
2) add 1 ml H2O. Dissolves easily.
3) divide into 0.2 ml portions
4) store frozen at -20 °C

Oxygraph-2k Manual Titrations: MiPNet09.12

  • Titration volume: 5 µl using a 25 µl syringe
  • Final conc. in 2 ml O2k-chamber: 10 µM