Difference between revisions of "ADP"
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::1) weigh 491.2 mg of ADP (1 mmol ADP) | ::1) weigh 491.2 mg of ADP (1 mmol ADP) | ||
::2) add 1.3 ml H<sub>2</sub>O | ::2) add 1.3 ml H<sub>2</sub>O | ||
::3) neutralize with 5 N KOH (approx. 465 µl) !ADP will dissolve after addition of KOH! (this is '''Solution A''') | ::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 | ::4) check pH and adjust to 7 if necessary | ||
| Line 33: | Line 33: | ||
'''MgCl<sub>2</sub>''' (Scharlau MA0036: MgCl<sub>2</sub>.6H<sub>2</sub>O, FW = 203.3) | '''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''' | ::1) add 121,98 mg MgCl<sub>2</sub> to '''Solution A''' !White precipitate will occur, which will be dissolved after 1-2 min of stirring on magnetic stirrer at room temperature! | ||
::2) follow steps 4-6 of the above instructions | ::2) follow steps 4-6 of the above instructions | ||
Revision as of 12:54, 3 December 2010
- high-resolution terminology - matching measurements at high-resolution
ADP
Description
Adenosine 5’diphosphate (D), C10H15N5O10P2K, potassium salt; substrate of ANT and ATP synthase.
Abbreviation: D
Reference: MiPNet03.02, MiPNet09.12
Labels:
Regulation: ATP; ADP; AMP; PCr"ATP; ADP; AMP; PCr" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property.
Application in HRR
D: ADP (Adenosine 5'diphosphate, C10H15N5O10P2K, potassium salt); Sigma A 5285, 1 g, store at -20 °C): FW = 491.2
Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.
Preparation of 500 mM ADP stock solution (dissolved in H2O):
- 1) weigh 491.2 mg of ADP (1 mmol ADP)
- 2) add 1.3 ml H2O
- 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 Mg2+:
To keep free [MgMg2+] constant during respiration measurement mix ADP (or ATP) with MgCl2 (0.6 mol/mol ADP or 0.8 mol/mol ATP).
MgCl2 (Scharlau MA0036: MgCl2.6H2O, FW = 203.3)
- 1) add 121,98 mg MgCl2 to Solution A !White precipitate will occur, which will be dissolved after 1-2 min of stirring on magnetic stirrer at room temperature!
- 2) follow steps 4-6 of the above instructions
- Comment: In some cases it was observed, that after defreezing of the 'ADP-Mg' a white precipitate has formed again. If this is the case we recommend to prepare the ADP stock solution and a MgCl2 solution (0.6 mol/mol ADP) seperately and perform a titration of both solutions in immediate succession.
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
