Difference between revisions of "ADP"

Revision as of 18:01, 19 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

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 (Sigma,A 5285: Adenosine 5'diphosphate, C₁₀H₁₅N₅O₁₀P₂K, potassium salt, FW=491.2)

Preparation of 500 mM ADP stock solution (dissolved in H₂O):

491.2 mg/2 ml H₂O of final volume (solution A)

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!

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 [Mg2+] 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, MW=203.3)

Solution A + MgCl₂

1-3) follow steps 1-3 of the above instructions

3b) add 121,98 mg MgCl₂

5-6) follow steps 5-6 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

@@ Line 10: / Line 10: @@
 |type=Substrate PS
 }}
-<h2>HRR</h2>
-<p>D: ADP (Sigma,A 5285: Adenosine 5&rsquo;diphosphate, C<sub>10</sub>H<sub>15</sub>N5O10P2K, potassium salt, MW=491.2)</p>
+== Application in [[HRR]] ==
-<p><strong>Preparation of stock solution</strong> (dissolved in H2O): </p>
-# 491.2 mg/2 ml of final volume. Weigh 491.2 mg of ADP, add 1 ml H2O and 450 ul 5 N KOH. ADP will dissolve after addition of KOH. Check pH and adjust to 7 (solution A).</p>
+'''D: ADP''' (Sigma,A 5285: 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, FW=491.2)
-# <strong>500 mM ADP stock solution</strong>: Adjust the total volume of solution A&nbsp;to 2 ml and divide into 0.2 ml portions. Store frozen at -80 &deg;C.</p>
-# <strong>500 mM </strong><strong>ADP stock solution with 3 mM free Mg2+</strong>: To keep free [Mg2+] constant at 3 mM when titrating the ADP stock solution into MiR06, mix ADP (solution A)&nbsp;with MgCl2 (0.6 mol/mol ADP). Adjusting pH of ADP to 7, add 121.98 mg of MgCl2 (Scharlau MA0036: MgCl2.6H2O, MW=203.3). White precipitate will occurr, which will be dissolved after 1-2 min of stirring on magnetic stirrer at room temperature. Adjust final volume to 2 ml, divide into 0.2 ml portions and store at -80 &deg;C.</p>
+'''Preparation of 500 mM ADP stock solution''' (dissolved in H<sub>2</sub>O):
-<p>Final conc. in 2 ml O2k-chamber: 2-5 mM; titrate 8-20 &micro;l using the 25 &micro;l syringe.  [[MiPNet09.12]]
+:: '''491.2 mg/2 ml H<sub>2</sub>O of final volume (solution A)'''
+::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!
+::3) neutralize with 5 N KOH (approx. 465 µl)  !ADP will dissolve after addition of KOH!
+::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 [Mg2+] 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: MgCl2.6H2O, MW=203.3)
+:: '''Solution A + MgCl<sub>2</sub>'''
+::1-3) follow steps 1-3 of the above instructions
+:: 3b) add 121,98 mg MgCl<sub>2</sub>
+::5-6) follow steps 5-6 of the above instructions
+Final conc. in 2 ml O2k-chamber: 2-5 mM; titrate 8-20 &micro;l using the 25 &micro;l syringe.  [[MiPNet09.12]]