Difference between revisions of "User talk:Spitzer Gudrun"

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


:::: '''As: Ascorbate sodium salt''' (Sodium L-ascorbate); Sigma A 7631, 100 g, store at RT; FW = 198.1.
::: '''Ama: Antimycin A''' (from ''Streptomyces sp.'') Sigma A 8674, 25 mg, store at -20 °C: FW = 540


:::: <span style="color:#8B008B"> '''Caution:''' Light sensitive (store solution in a dark glass vial).</span>
:::: <span style="color:#8B008B"> '''Caution:''' '''Very toxic.''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.</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
::::# 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''':  
::: '''Preparation of 5 mM stock solution''' (dissolved in 99.9% ethanol) for '''2-mL O2k-chamber''':


::::# Weigh 1.980 g of ascorbate sodium salt and dissolve in 10 mL H<sub>2</sub>O
::::# Weigh 5.4 mg of antimycin A into a small glass vial. Caution: Ama is hygroscopic.
::::# To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.mL<sup>-1</sup> solution of pH 2).  
::::# Add 2 mL ethanol.
::::# Divide into 0.2 mL portions.  
::::# Divide into 0.2 mL portions in clear glass vials.
::::# Store frozen at -20 °C.
::::# Store at -20 °C.


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


:::»'''O2k manual titrations:''' [[MiPNet09.12]]
::::* Titration volume ('''2-mL O2k-chamber'''): 1 µL using a 10 µL Hamilton syringe.
::::* Final concentration: 2.5 µM.


::::* Titration volume ('''2-mL O2k-chamber'''): 5 µL of 800mM stock solution using a 25 µL Hamilton syringe.
::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL of 1000mM stock solution using a 10 µL Hamilton syringe
::::* Final concentration: 2 mM.


== Asc gutest 2 ==
::: '''Preparation of 1,25 mM stock solution''' (dissolved in 99.9% ethanol) for '''0.5-mL O2k-chamber''':


:::: '''As: Ascorbate sodium salt''' (Sodium L-ascorbate); Sigma A 7631, 100 g, store at RT; FW = 198.1.
::::# Take 100 µL of 5 mM stock solution.
 
::::# Add 300 µL ethanol.
:::: <span style="color:#8B008B"> '''Caution:''' Light sensitive (store solution in a dark glass vial).</span>
:::» '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]]
 
 
::: '''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
::::# 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 ('''2-mL O2k-chamber'''): 5 µL of 800 mM stock solution using a 25 µL Hamilton syringe.
::::* Final concentration: 2 mM.
 
 
::: '''Preparation of 1000 mM stock solution''' (dissolved in H<sub>2</sub>O) for use in '''0.5-mL O2k-chamber''':
 
::::# Weigh 1.980 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]]
 
::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL of 1000 mM stock solution using a 10 µL Hamilton syringe.
::::* Final concentration: 2 mM.
 
== Version 3 ==
 
:::: '''As: Ascorbate sodium salt''' (Sodium L-ascorbate); Sigma A 7631, 100 g, store at RT; FW = 198.1.
 
:::: <span style="color:#8B008B"> '''Caution:''' Light sensitive (store solution in a dark glass vial).</span>


::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL using a 10 µL Hamilton syringe.
::::* Final concentration: 2.5 µM.


:::: '''Preparation of 800 mM stock solution''' (dissolved in H<sub>2</sub>O) for '''2-mL O2k-chamber''':
== Antimycin A version 2 ==


::::# Weigh 1.584 g of ascorbate sodium salt and dissolve in 10 mL H<sub>2</sub>O
::: '''Ama: Antimycin A''' (from ''Streptomyces sp.'') Sigma A 8674, 25 mg, store at -20 °C: FW = 540
::::# 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]]
:::: <span style="color:#8B008B"> '''Caution:''' '''Very toxic.''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.</span>
 
::::* Titration volume ('''2-mL O2k-chamber'''): 5 µL using a 25 µL Hamilton syringe.
::::* Final concentration: 2 mM.
 
 
:::: '''Preparation of 1000 mM stock solution''' (dissolved in H<sub>2</sub>O) for '''0.5-mL O2k-chamber''':
 
::::# Weigh 1.980 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]]
 
::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL using a 10 µL Hamilton syringe.
::::* Final concentration: 2 mM.


== O2k manual titrations - list of chemicals ==


=== same conc for both chambers ===
::: '''Preparation of 5 mM stock solution''' (dissolved in 99.9% ethanol):


:::: This text is a test
::::# Weigh 5.4 mg of antimycin A into a small glass vial. Caution: Ama is hygroscopic.
:::: And even more words
::::# 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.


=== diff conc for sV and lV chamber ===
:::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]


::::* Another test
::::* Titration volume ('''2-mL O2k-chamber'''): 1 µL (5 mM) using a 10 µL Hamilton syringe.
::::* Third line of the test
::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL (1.25 mM) using a 10 µL Hamilton syringe.
::::* Final concentration: 2.5 µM.

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.