Difference between revisions of "ADP"
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== Application in [[HRR]] == | == Application in [[HRR]] == | ||
'''D: ADP''' ( | '''D: ADP''' (Adenosine 5'diphosphate, C<sub>10</sub>H<sub>15</sub>N<sub>5</sub>O<sub>10</sub>P<sub>2</sub>K, potassium salt; Sigma A 5285 (-20 °C): FW = 491.2) | ||
'''Preparation of 500 mM ADP stock solution''' (dissolved in H<sub>2</sub>O): | '''Preparation of 500 mM ADP stock solution''' (dissolved in H<sub>2</sub>O): | ||
::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 !White precipitate will occur, which will be dissolved after 1-2 min of stirring on magnetic stirrer at room temperature! | ::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! | ::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 | ||
::5) adjust final volume to 2 ml and divide into 0.2 ml portions. | ::5) adjust final volume to 2 ml and divide into 0.2 ml portions. | ||
::6) store frozen at -80 °C | ::6) store frozen at -80 °C | ||
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'''Preparation of 500 mM ADP stock solution with 3 mM free | '''Preparation of 500 mM ADP stock solution with 3 mM free Mg<sup>2+</sup>''': | ||
To keep free [ | 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>''' (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 | ::2) follow steps 4-6 of the above instructions | ||
:: | |||
Final conc. in 2 ml O2k-chamber: 2-5 mM; titrate 8-20 µl using the 25 µl syringe. [[MiPNet09.12]] | Final conc. in 2 ml O2k-chamber: 2-5 mM; titrate 8-20 µl using the 25 µl syringe. [[MiPNet09.12]] | ||
Revision as of 10:17, 22 November 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 (-20 °C): FW = 491.2)
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 !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 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
- 2) follow steps 4-6 of the above instructions
Final conc. in 2 ml O2k-chamber: 2-5 mM; titrate 8-20 µl using the 25 µl syringe. MiPNet09.12
