in vitro PHARMACOLOGY 2011 CATALOG - Cerep

126 in vitro pharmacology 2011 catalog
❚ protein-SERINE/THREONINE kinases [CMGC]
CLK2
Source
human recombinant (insect cells)
Substrate
ATP + biotinyl-βAβAβAAGAGKRREILSRRPS
YRK (10 nM)
Measured product phospho-biotinyl-βAβAβAAGAGKRREILSRR
Ref. 2019
PSYRK
Detection method HTRF
Q 3 weeks
Reference
Ro-318220 (IC
Included in:
50 : 75 nM)
Comprehensive kinase profile Nayler, O. et al. (1998) J. Biol. Chem., 273: 34341-34348.
enzyme activity (% of control)
100
50
0
Ro-318220
staurosporine
K252a
H-89
-10 -9 -8 -7 -6 -5 -4 -3
log [drug] (M)
DYRK1a
Ref. 2781
Q 3 weeks
Included in:
Organ safety profile
Comprehensive kinase profile
Source
human recombinant
Substrate
ATP + Ulight-CFFKNIVTPRTPPPSQGK-amide
(100 nM)
Measured product phospho-Ulight-CFFKNIVTPRTPPPSQGKamide
Detection method LANCE
Reference
staurosporine (IC 50 : 16 nM)
Himpel, S. et al. (2000) J. Biol. Chem., 275: 2431-2438.
-12 -11 -10 -9 -8 -7 -6 -5 -4
-10 -9 -8 -7 -6
-10 -7 -9 -8 -6 -5 -4
-11 -10 -6 -5
-12 -9 -8 -7
DYRK2
Ref. 2788
Q 3 weeks
Included in:
Comprehensive kinase profile
Source
human recombinant
Substrate
ATP + Ulight-CFFKNIVTPRTPPPSQGK-amide
(100 nM)
Measured product phospho-Ulight-CFFKNIVTPRTPPPSQGKamide
Detection method LANCE
Reference
staurosporine (IC 50 : 1100 nM)
Taira, N. et al. (2007) Mol. Cell., 25: 725-738.
DYRK3
Ref. 1963
Q 3 weeks
Source
human recombinant (insect cells)
Substrate
ATP + biotinyl-MBP
(40 nM)
Measured product phospho-biotinyl-MBP
Detection method HTRF
Reference
staurosporine (IC 50 : 37 nM)
Li, K. et al. (2002) J. Biol. Chem., 277: 47052-47060.
enzyme activity (% of control)
100
50
staurosporine
Ro-318220
K252a
0
H-89
-8 -7 -6 -4
-9 -5
log [drug] (M)
DYRK4
Ref. 1962
Q 3 weeks
Included in:
Comprehensive kinase profile
Source
human recombinant (insect cells)
Substrate
ATP + biotinyl-MBP
(80 nM)
Measured product phospho-biotinyl-MBP
Detection method HTRF
Reference
hymenialdisine (IC 50 : 149 nM)
Li, K. et al. (2002) J. Biol. Chem., 277: 47052-47060.
enzyme activity (% of control)
-10 -9 -8 -7 -6 -5 -4
100
-9 -8 -7 -6 -5 -4
50
0
-10 -9 -5 -8 -7 -6 -4
log [drug] (M)
hymenialdisine
staurosporine
Bis 10
EGCG
❚ Assays converted from HTRF ® to LANCEUltra ® technology
A majority of our kinase assays have been converted from HTRF ® to LANCEUltra ® technology. Both technologies are TR-FRET with the
difference in europium labelling: europium cryptate for HTRF ® and europium chelate for LANCEUltra ® .
Assays are converted following a standard procedure consisting of time course experiment, Km determination and pharmacology
characterization of the enzyme by inhibiting its activity with known inhibitors. IC 50 values obtained for known inhibitors are compared
to literature and to those obtained using HTRF ® technology. If all values are in agreement with the previous ones, LANCE ® technology is
validated for the kinase of interest.
This conversion allows us to standardize and automate our kinase assays providing shorter turnaround times for both profiling and screening
experiments.