in vitro PHARMACOLOGY 2011 CATALOG - Cerep

138 in vitro pharmacology 2011 catalog
❚ protein-SERINE/THREONINE kinases [AGC]
GRK2 (ADRBK1)
Ref. 2200
Q 3 weeks
Included in:
Organ safety profile
Comprehensive kinase profile
Source
human recombinant (insect cells)
Substrate ATP + His-tagged Rb truncated protein
(140 nM)
Measured product phospho-His-tagged Rb truncated protein
Detection method HTRF
Reference
staurosporine (IC 50 : 50 nM)
Laccarino, G. et al. (2005) Eur. Heart. J., 26: 1752-1758.
GRK3/BARK2 (ADRBK2)
Ref. 2627
Q 3 weeks
Included in:
Organ safety profile
Comprehensive kinase profile
Source
human recombinant
Substrate ATP + Ulight-ARTKQTARKSTGGKAPRKQLAGCG
(25 nM)
Measured product phospho-Ulight-ARTKQTARKSTGGKAPRKQL
AGCG
Detection method LANCE
Reference
staurosporine (IC 50 : 295 nM)
Luo, J. and Benovic, J.L. (2003) J. Biol. Chem., 278: 50908-50914.
GRK5
Ref. 2020
Q 3 weeks
Source
human recombinant (insect cells)
Substrate ATP + biotinyl-KKKKERLLDDRHDSGLDSMKDEE
(600 nM)
Measured product phospho-biotinyl-KKKKERLLDDRHDSGLDSKDEE
Detection method HTRF
Reference
staurosporine (IC 50 : 27 nM)
Kunapuli, P. et al. (1994) J. Biol. Chem., 269: 1099-1106.
MRCKa
Ref. 2629
Q 3 weeks
Included in:
Comprehensive kinase profile
Source
human recombinant
Substrate ATP + Ulight-ARTKQTARKSTGGKAPRKQLAGCG
(75 nM)
Measured product phospho-Ulight-ARTKQTARKSTGGKAPRKQL
AGCG
Detection method LANCE
Reference staurosporine (IC 50 : 18 nM)
Tan, I. et al. (2001) Mol. Cell. Biol., 21: 2767-2778.
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.