Determination of ATP and ADP in blood platelets: A modification of the firefly luciferase assay for plasma

✍ Scribed by Holm Holmsen; Eva Storm; H. James Day

• Publisher: Elsevier Science
• Year: 1972
• Tongue: English
• Weight: 719 KB
• Volume: 46
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(72)90323-5

✦ Synopsis

EDTA, pH 7.4 (Na,H,EDTA adjusted with NaOH or Na,EDTA adjusted with. HCl). EDTA is not completely soluble in this mixture, and the precipitated solid becomes finely dispersed in fresh mixtures. Large clumps form after 30 min of storage and E-E was therefore used fresh. On mixing E-E with sample (or standard) the finely dispersed EDTA particles dissolved instantaneously.

Platelet-rich plasma (PRP) was prepared from fresh human blood (9 vol blood + 1 vol 0.11 M Na, citrate) by centrifugation at room temperature for 15 min at G,,,, = 3209 or for 3 min at G,,,, = 14359. The supernatant (PRP) was saved. Ocassionally, the remainder of the blood was centrifuged for 30 min at G,,, = 3500g in order to obtain platelet-poor plasma (PPP) .

Platelet suspension. Platelets were isolated by centrifugation at G,,,, = 17,000g at 4" for 10 min and resuspended in the medium described by Haslam ( 5)) with the EDTA concentration increased to 4.8 mM.