in vitro PHARMACOLOGY 2011 CATALOG - Cerep

148 in vitro pharmacology 2011 catalog
❚ protein-SERINE/THREONINE kinases [STE]
PAK1
Ref. 1934
Q 3 weeks
Included in:
Organ safety profile
Comprehensive kinase profile
Source
human recombinant (insect cells)
Substrate ATP + biotinyl-tyrosine hydroxylase-derived
peptide (40 nM)
Measured product phospho-biotinyl-tyrosine hydroxylasederived
peptide
Detection method HTRF
Reference
staurosporine (IC 50 : 9 nM)
Yang, Z. et al. (2004) Clin. Cancer. Res., 10: 658-667.
enzyme activity (% of control)
100
50
0
-11 -10 -9 -8 -7 -6 -5 -4
log [drug] (M)
staurosporine
Ro-318220
K252a
H-89
-3
PAK2
Ref. 2932
Q 3 weeks
Included in:
ExpresS Diversity kinase profile
Comprehensive kinase profile
Source
human recombinant (Sf9 cells)
Substrate ATP + Ulight-RRRSLLE
(50 nM)
Measured product phospho-Ulight-RRRSLLE
Detection method LANCE
Reference
-10
staurosporine (IC 50 : 21 nM)
Wu, H. and Wang, Z-X. (2003) J. Biol. Chem., 278: 41768-41778.
-9 -8 -7 -6 -5 -4
-10 -9 -8 -7 -6 -5 -4 -3
-8 -7 -6 -5 -4
-10 -9 -8 -7 -6 -5
-9 -8 -7 -6 -5 -4
-10 -9 -8 -7 -6 -5
PAK4
Ref. 3363
Q 3 weeks
Included in:
ExpresS Diversity kinase profile
Comprehensive kinase profile
Source
human recombinant (insect cells)
Substrate ATP + Ulight-RRRSLLE
(50 nM)
Measured product phospho-Ulight-RRRSLLE
Detection method LANCE
Reference
staurosporine (IC 50 : 77.5 nM)
Arias-Romero, L.E. and Chernoff, J. (2008) Biol. Cell., 100: 97-108.
TAOK2 (TAO1)
Ref. 2586
Q 3 weeks
Included in:
ExpresS Diversity kinase profile
Organ safety profile
Comprehensive kinase profile
Source
human recombinant
Substrate ATP + Ulight-FLGFTYVAP
(40 nM)
Measured product phospho-Ulight-FLGFTYVAP
Detection method LANCE
Reference
staurosporine (IC 50 : 23 nM)
Hutchison, M. et al. (1998) J. Biol. Chem., 273: 28625-28632.
selected cerep assays
❚ Biochemical kinase assays
Kinase assays are enzymatic assays mostly using activated kinases. Those are usually full length kinase or cytoplasmic domain of RTK.
Assays are designed to be as close as possible to ATP and substrate Km.
The technology used to measure substrate phosphorylation is TR-FRET (HTRF ® or LANCE ® ). A few assays are cascade of activation.
❚ Cellular kinase assays
In order to complement our existing biochemical assay platform, we have implemented cellular kinase assays. These assays allow to
confirm inhibitors activity in a relevant cellular background and profile their selectivity against multiple signaling pathways.
- Cellular kinase phosphorylation assays
Cellular kinase phosphorylation assays are developed using AlphaScreen ® Surefire ® assay kits. The assays are optimized for directly
measuring kinase activation following treatment of cells with activators of signaling pathways.
- Cellular tyrosine kinase receptor activity assays
We have demonstrated that the impedance-based technology can be used to monitor the activity of the main tyrosine kinase receptor
families. These label-free assays allow the identification of inhibitors targeting either the ligand binding domain or the kinase domain.
These assays can be developed in virtually any cancer cell lines or primary cells. As an example, we have developed and validated
the EGFR cellular assay in the following cancer cell lines: A431, HELA and MDA-MB-231.
❚ Binding kinase assays
See page 107.