in vitro PHARMACOLOGY 2011 CATALOG - Cerep

132 in vitro pharmacology 2011 catalog
❚ protein-SERINE/THREONINE kinases [CaMK]
DAPK1
Ref. 1717
Q 3 weeks
Included in:
Organ safety profile
Comprehensive kinase profile
Source
human recombinant (insect cells)
Substrate ATP + biotinyl-βAβAβAKKLNRTLSFAEP
(800 nM)
Measured product phospho-biotinyl-βAβAβAKKLNRTLSFAEP
Detection method HTRF
Reference
staurosporine (IC 50 : 80 nM)
Valentza, A.V. et al. (2001) J. Biol. Chem., 276: 38956-38965.
enzyme activity (% of control)
100
50
0
-9 -8 -7 -6 -5 -4
log [drug] (M)
staurosporine
H-89
Ro-318220
K252a
DAPK2
Ref. 1930
Q 3 weeks
Included in:
Organ safety profile
Comprehensive kinase profile
Source
human recombinant (insect cells)
Substrate ATP + biotinyl-βAβAβAKKLNRTLSFAEP
(200 nM)
Measured product phospho-biotinyl-βAβAβAKKLNRTLSFAEP
Detection method HTRF
Reference
staurosporine (IC 50 : 8.3 nM)
Kavai, T. et al. (1999) Oncogene, 18: 3471-3480.
enzyme activity (% of control)
-10 -9 -8 -7 -6 -5 -4
100
-8 -7 -6 -5 -4
50
-10 -9 -8 -7 -6 -5 -4
staurosporine
Ro-318220
H-89
0
K252a
-10 -9 -8 -7 -6 -5 -4
log [drug] (M)
DCAMKL1
Ref. 2613
Q 3 weeks
Included in:
Organ safety profile
Comprehensive kinase profile
Source
human recombinant
Substrate ATP + CREBtide (CKRREILSRRPSYRK)
(25 nM)
Measured product phospho-CREBtide (CKRREILSRRPSYRK)
Detection method LANCE
Reference
staurosporine (IC 50 : 80 nM)
Lin, P.T. et al. (2000) J. Neurosc., 20: 9152-9161.
-9 -8 -7 -6 -5 -4
-8 -7 -6 -5 -4
-10 -9 -8 -7 -6 -5 -4
DCAMKL2
Ref. 2741
Q 3 weeks
Included in:
Comprehensive kinase profile
Source
human recombinant
Substrate ATP + CREBtide (CKRREILSRRPSYRK)
(25 nM)
Measured product phospho-CREBtide (CKRREILSRRPSYRK)
Detection method LANCE
Reference
staurosporine (IC 50 : 14.5 nM)
Ohmae, S. et. al. (2006) J. Biol. Chem., 281: 20427-20439.
DRAK1
Ref. 2930
Q 3 weeks
Included in:
Comprehensive kinase profile
Source
human recombinant
Substrate ATP + Ulight-ARTKQTARKSTGGKAPRKQLA
GCG (50 nM)
Measured product phospho-Ulight-ARTKQTARKSTGGKAPRKQ
LAGCG
Detection method LANCE
Reference staurosporine (IC 50 : 48 nM)
Sanjo, H. et al. (1998) J. Biol. Chem, 273: 29066-29071.
❚ 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.