ADP

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 adjusted to 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 !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

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