User talk:Spitzer Gudrun: Difference between revisions

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= Asc gutest 1=
== Antimycin A version 1 ==


== Application in [[HRR]] ==
::: '''Ama: Antimycin A''' (from ''Streptomyces sp.'') Sigma A 8674, 25 mg, store at -20 °C: FW = 540


:::: '''As: Ascorbate sodium salt''' (Sodium L-ascorbate); Sigma A 7631, 100 g, store at RT; FW = 198.1.
:::: <span style="color:#8B008B"> '''Caution:''' '''Very toxic.''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.</span>


:::: <span style="color:#8B008B"> '''Caution:''' Light sensitive (store solution in a dark glass vial).</span>
'
::: '''Preparation of 800 mM stock solution''' (dissolved in H<sub>2</sub>O) for use in '''2-mL O2k-chamber''':


::::# Weigh 1.584 g of ascorbate sodium salt and dissolve in 10 mL H<sub>2</sub>O
::: '''Preparation of 5 mM stock solution''' (dissolved in 99.9% ethanol) for '''2-mL O2k-chamber''':
::::# To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.mL<sup>-1</sup> solution of pH 2).
::::# Divide into 0.2 mL portions.
::::# Store frozen at -20 °C.


::: '''Preparation of 1000 mM stock solution''' (dissolved in H<sub>2</sub>O) for use in '''0.5-mL O2k-chamber''':
::::# Weigh 5.4 mg of antimycin A into a small glass vial. Caution: Ama is hygroscopic.
::::# Add 2 mL ethanol.
::::# Divide into 0.2 mL portions in clear glass vials.
::::# Store at -20 °C.


::::# Weigh 1.980 g of ascorbate sodium salt and dissolve in 10 mL H<sub>2</sub>O
:::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
::::# To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.mL<sup>-1</sup> solution of pH 2).
::::# Divide into 0.2 mL portions.
::::# Store frozen at -20 °C.


::::* Titration volume ('''2-mL O2k-chamber'''): 1 µL using a 10 µL Hamilton syringe.
::::* Final concentration: 2.5 µM.


:::»'''O2k manual titrations:''' [[MiPNet09.12]]


::::* Titration volume ('''2-mL O2k-chamber'''): 5 µL 800mM using a 25 µL Hamilton syringe.
::: '''Preparation of 1,25 mM stock solution''' (dissolved in 99.9% ethanol) for '''0.5-mL O2k-chamber''':
::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL 1000mM using a 10 µL Hamilton syringe
::::* Final concentration: 2 mM.
= Asc gutest 2 =


== Application in [[HRR]] ==
::::# Take 100 µL of 5 mM stock solution.
::::# Add 300 µL ethanol.
:::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]


:::: '''As: Ascorbate sodium salt''' (Sodium L-ascorbate); Sigma A 7631, 100 g, store at RT; FW = 198.1.
::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL using a 10 µL Hamilton syringe.
::::* Final concentration: 2.5 µM.


:::: <span style="color:#8B008B"> '''Caution:''' Light sensitive (store solution in a dark glass vial).</span>
== Antimycin A version 2 ==


::: '''Ama: Antimycin A''' (from ''Streptomyces sp.'') Sigma A 8674, 25 mg, store at -20 °C: FW = 540


::: '''Preparation of 800 mM stock solution''' (dissolved in H<sub>2</sub>O) for '''2-mL O2k-chamber''':
:::: <span style="color:#8B008B"> '''Caution:''' '''Very toxic.''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.</span>


::::# Weigh 1.584 g of ascorbate sodium salt and dissolve in 10 mL H<sub>2</sub>O
::::# To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.mL<sup>-1</sup> solution of pH 2).
::::# Divide into 0.2 mL portions.
::::# Store frozen at -20 °C.


:::»'''O2k manual titrations:''' [[MiPNet09.12]]
::: '''Preparation of 5 mM stock solution''' (dissolved in 99.9% ethanol):


::::* Titration volume ('''2-mL O2k-chamber'''): 5 µL 800 mM using a 25 µL Hamilton syringe.
::::# Weigh 5.4 mg of antimycin A into a small glass vial. Caution: Ama is hygroscopic.
::::* Final concentration: 2 mM.
::::# Add 2 mL ethanol.
::::# Divide into 0.2 mL portions in clear glass vials.
::::# Store at -20 °C.
::::# For '''1.25''' mM concentration take 100 µL stock solution (5 mM) and dilute it with 300 µl ethanol.


:::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]


::: '''Preparation of 1000 mM stock solution''' (dissolved in H<sub>2</sub>O) for '''0.5-mL O2k-chamber''':  
::::* Titration volume ('''2-mL O2k-chamber'''): 1 µL (5 mM) using a 10 µL Hamilton syringe.
 
::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL (1.25 mM) using a 10 µL Hamilton syringe.
::::# Weigh 1.980 g of ascorbate sodium salt and dissolve in 10 mL H<sub>2</sub>O
::::* Final concentration: 2.5 µM.
::::# To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.mL<sup>-1</sup> solution of pH 2).
::::# Divide into 0.2 mL portions.
::::# Store frozen at -20 °C.
 
:::»'''O2k manual titrations:''' [[MiPNet09.12]]
 
::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL 1000 mM using a 10 µL Hamilton syringe.
::::* Final concentration: 2 mM.

Latest revision as of 12:12, 23 October 2020

Antimycin A version 1

Ama: Antimycin A (from Streptomyces sp.) Sigma A 8674, 25 mg, store at -20 °C: FW = 540
Caution: Very toxic. Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.


Preparation of 5 mM stock solution (dissolved in 99.9% ethanol) for 2-mL O2k-chamber:
  1. Weigh 5.4 mg of antimycin A into a small glass vial. Caution: Ama is hygroscopic.
  2. Add 2 mL ethanol.
  3. Divide into 0.2 mL portions in clear glass vials.
  4. Store at -20 °C.
» O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (2-mL O2k-chamber): 1 µL using a 10 µL Hamilton syringe.
  • Final concentration: 2.5 µM.


Preparation of 1,25 mM stock solution (dissolved in 99.9% ethanol) for 0.5-mL O2k-chamber:
  1. Take 100 µL of 5 mM stock solution.
  2. Add 300 µL ethanol.
» O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (0.5-mL O2k-chamber): 1 µL using a 10 µL Hamilton syringe.
  • Final concentration: 2.5 µM.

Antimycin A version 2

Ama: Antimycin A (from Streptomyces sp.) Sigma A 8674, 25 mg, store at -20 °C: FW = 540
Caution: Very toxic. Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.


Preparation of 5 mM stock solution (dissolved in 99.9% ethanol):
  1. Weigh 5.4 mg of antimycin A into a small glass vial. Caution: Ama is hygroscopic.
  2. Add 2 mL ethanol.
  3. Divide into 0.2 mL portions in clear glass vials.
  4. Store at -20 °C.
  5. For 1.25 mM concentration take 100 µL stock solution (5 mM) and dilute it with 300 µl ethanol.
» O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (2-mL O2k-chamber): 1 µL (5 mM) using a 10 µL Hamilton syringe.
  • Titration volume (0.5-mL O2k-chamber): 1 µL (1.25 mM) using a 10 µL Hamilton syringe.
  • Final concentration: 2.5 µM.
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