Coupling Rates for 20 Minute Kinase Reactions

The following table lists the coupling rates for 20 minute kinase reactions coupled with Recombinant Mouse CD39L2.  The coupling rates are for 50 μL reactions carried out at room temperature in a buffer consisting of 25 mM HEPES, 10 mM CaCl2, 10 mM MgCl2, 150 mM NaCl, pH 7.0. The coupling rates were calculated using a specific rate constant of 97.78 nmol/min/μg/mM.

The red shaded region represents the optimal range for coupling rates in order to achieve the maximum signal-to-noise ratio for the phosphatase-coupled kinase reaction.

For more detailed technical information, please also see: Wu, Z.L. (2011) PLoS ONE 6(8):e23172.

Coupling Rates for 20 Minute Kinase Reactions

ATP
(μM)		 CD39L2 (μg)
0.025 0.050 0.075 0.100 0.125 0.150 0.175 0.200
50 0.323 0.515 0.634 0.712 0.765 0.802 0.830 0.851
100 0.309 0.498 0.618 0.697 0.752 0.791 0.820 0.842
150 0.299 0.486 0.606 0.686 0.742 0.783 0.813 0.836
200 0.291 0.475 0.596 0.677 0.734 0.776 0.806 0.830
250 0.285 0.467 0.587 0.669 0.727 0.769 0.801 0.825
300 0.279 0.460 0.580 0.662 0.721 0.764 0.796 0.821
400 0.269 0.447 0.567 0.650 0.710 0.754 0.787 0.813
500 0.262 0.436 0.556 0.640 0.700 0.745 0.779 0.806
600 0.255 0.427 0.546 0.631 0.692 0.738 0.773 0.800
700 0.249 0.419 0.537 0.622 0.684 0.731 0.766 0.794
800 0.244 0.411 0.530 0.615 0.677 0.724 0.761 0.789
900 0.239 0.405 0.522 0.608 0.671 0.718 0.755 0.784
1000 0.234 0.399 0.516 0.601 0.665 0.713 0.750 0.779
1100 0.230 0.393 0.510 0.595 0.659 0.708 0.745 0.775
1200 0.227 0.388 0.504 0.590 0.654 0.703 0.741 0.771
1300 0.223 0.383 0.499 0.584 0.649 0.698 0.736 0.767
1400 0.220 0.378 0.493 0.579 0.644 0.693 0.732 0.763
1500 0.217 0.374 0.489 0.574 0.639 0.689 0.728 0.760
1600 0.214 0.370 0.484 0.570 0.635 0.685 0.725 0.756
1700 0.212 0.366 0.480 0.565 0.631 0.681 0.721 0.753
1800 0.209 0.362 0.476 0.561 0.626 0.677 0.717 0.749
1900 0.207 0.358 0.472 0.557 0.623 0.674 0.714 0.746
2000 0.204 0.355 0.468 0.553 0.619 0.670 0.711 0.743
2100 0.202 0.352 0.464 0.549 0.615 0.667 0.707 0.740
2200 0.200 0.349 0.460 0.546 0.612 0.663 0.704 0.737
2300 0.198 0.346 0.457 0.542 0.608 0.660 0.701 0.735
2400 0.196 0.343 0.454 0.539 0.605 0.657 0.698 0.732
2500 0.194 0.340 0.451 0.536 0.602 0.654 0.696 0.729
2600 0.193 0.337 0.448 0.532 0.599 0.651 0.693 0.727
2700 0.191 0.335 0.445 0.529 0.596 0.648 0.690 0.724
2800 0.189 0.332 0.442 0.526 0.593 0.645 0.687 0.722
2900 0.188 0.330 0.439 0.523 0.590 0.642 0.685 0.719
3000 0.186 0.327 0.436 0.521 0.587 0.640 0.682 0.717