in vitro
pharmacology
kinase assays
Cerep’s panel of kinases has increased from 205 biochemical assays as of January,1, 2009 to 270 (including
16 cellular kinase assays) as of January 1, 2010 allowing to provide a wider coverage of all major families in the
human kinome (Manning G. et al. (2002) Science, 298: 1912-1934).
Cerep kinase assays

Biochemical kinase assays
Majority of our biochemical kinase assays use activated kinases. They 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.
- Assays converted from HTRF® to LANCEUltra® technology
A number of our kinase assays has 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. IC50 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.
NOTE: The converted assays are indicated  . They supersede the assays previously listed in our catalog.

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.
 To expand our existing kinase services, we offer binding kinase assays to identify type II inhibitors on unphosphorylated kinases, to
compare compounds activity on unphosphorylated and phophorylated kinases and to discriminate between ATP and non-ATP competitive
inhibitors. In addition, we are now using ADP readout to develop kinase assays on intractable substrates (for which no antibody exist)
which allows us to profile kinase activity on different natural substrates in order to identify substrate selective inhibitors. These assays are
displayed as part of our custom research. For further information, please contact us at customresearch@cerep.com.
protein-tyrosine kinases ❚
Family/assay
protein-tyrosine kinases / RTK
ALK
 Axl kinase
 c-kit kinase
 c-Met kinase
c  c-Met kinase (HGFR)
 DDR2 kinase
 EGFR kinase
c  EGFR kinase (Hela cells)
c  EGFR kinase (A431 cells) - impedance
c  EGFR kinase (A431 cells) - AlphaScreen
c  EGFR kinase (MDA-MB-231 cells)
EphA1 kinase
 EphA2 kinase
 EphA3 kinase
EphA4 kinase
EphA5 kinase
 EphA6 kinase
 EphA7 kinase
c
cellular assay

new assay
sales@cerep.com ❚ www.cerep.com

new protocol

Ref.
➁
➁
➁
➀ ➁
➁
➀ ➁
➁
➀ ➁
➀ ➁
➁
➁
➁
➁
2678
3062
3070
2867
G150
2783
2865
G160
G161
G151
G162
1873
3055
3066
1702
2640
3071
3058
Family/assay
protein-tyrosine kinases / RTK (cont'd)
 EphA8 kinase
 EphB1 kinase
 EphB2 kinase
 EphB3 kinase
 EphB4 kinase
 FGFR1 kinase
 FGFR2 kinase
 FGFR3 kinase
 FGFR4 kinase
c  FGFR kinase
 FLT-1 kinase (VEGFR1)
 FLT-3 kinase
 FLT-4 kinase (VEGFR3)
 Fms/CSFR kinase
HER2/ErbB2 kinase
HER4/ErbB4 kinase
 IGF1R kinase
c  IGF1R kinase
assay converted from HTRF® to LANCEUltra®
Ref.
➀
➀
➀
➀
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
3072
3053
3054
2731
3059
2868
2893
2894
2895
G152
3068
2866
2896
3069
1598
1875
3061
G163
human ➀ ExpresS Diversity kinase profile ➁ Comprehensive kinase profile
ASSAY LIST

January 2010
kinase ASSAY LIST

January 2010
protein-tyrosine kinases ❚ (cont'd)
Family/assay
protein-tyrosine kinases / RTK (cont'd)
 IRK (InsR)
c  IRK (InsRb)
 IRR kinase
 KDR kinase (VEGFR2)
LTK
Mer kinase
 MusK
 PDGFRa kinase
 PDGFRb kinase
c  PDGFR kinase
Ret kinase
 Ron kinase
Ros kinase
 TIE2 kinase
 TRKA
 TRKB
 TRKC
 Tyro3/Sky kinase
c  VEGFR kinase
protein-tyrosine kinases / CTK
 Abl kinase
 Ack
 Arg kinase
BLK
BMX (Etk) kinase
 Brk
BTK
Ref.
➀ ➁
➁
➀ ➁
➁
➁
➁
➁
➁
➁
➁
➁
➀ ➁
➁
➁
➁
➀ ➁
➁
➁
➁
➁
➁
2898
G153
3073
2864
2642
2643
2899
3064
2900
G154
1593
3067
1594
2736
2901
2902
3080
3081
G142
3056
3052
3057
1697
1587
3063
1589
Family/assay
protein-tyrosine kinases / CTK (cont'd)
CSK
CTK
 FAK
Fer kinase
Fes kinase
 Fgr kinase
 FRK
Fyn kinase
HCK
 ITK
JAK1
 JAK2
 JAK3
 Lck kinase
 Lyn A kinase
Lyn B kinase
 PYK2
 Src kinase
Srm kinase
 Syk
TEC kinase
 Tnk1
 TXK
Tyk2 (JTK1)
Yes kinase
ZAP70 kinase
Ref.
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➀ ➁
➀ ➁
➁
➁
➁
➀ ➁
➁
➁
➁
➁
➁
➁
➁
1524
2638
3065
2641
1519
3060
2784
0212
1590
2733
2619
2869
2905
2906
0669
2201
3077
2907
2646
2780
2647
2785
2786
2621
1595
0705
protein-serine/threonine kinases ❚
Family/assay
protein-serine/threonine kinases / CMGC
 CDC2/CDK1 (cycB)
 CDK2 (cycA)
CDK3 (cycE1)
CDK4 (cycD1)
 CDK5/p35
 CDK6 (cycD3)
 CDK7/MAT1 (cycH)
 CDK8 (cycC)
 CDK9 (cycT1)
 CK2 (casein kinase 2)
 CLK1
CLK2
 DYRK1a
 DYRK2
DYRK3
DYRK4
 ERK
1
 ERK (P42mapk)
2
c  ERK1/2
ERK5 (MAPK7)
 GSK3a
 GSK3b
c  GSK3b
 HIPK2
HIPK3
HIPK4
 JNK1
JNK2
 JNK3
c  JNK1/3
 p38a kinase
p38b2 kinase (SAPK2b2)
p38g kinase
 p38d kinase
PCTAIRE1 kinase
2
Ref.
➀ ➁
➀ ➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➀ ➁
➁
➁
➀ ➁
➁
➁
➀ ➁
➁
➁
➀ ➁
➁
➁
➁
2875
2908
2679
0796
2877
2909
2910
2911
2912
2913
2787
2019
2781
2788
1963
1962
2914
2878
G155
2680
2842
2879
G156
2915
1980
1981
2880
1470
2916
G157
2881
1960
1580
3116
2025
Family/assay
protein-serine/threonine kinases / CaMK
AMPKa
 BRSK1
 CaMK1a
 CaMK1d
 CaMK2a
 CaMK2g
CaMK4
 CHK1
 CHK2
DAPK1
DAPK2
DCAMKL1
 DCAMKL2
 DRAK1
MAPKAPK2
MAPKAPK3
 MAPKAPK5 (PRAK)
MARK1
MARK2
 MARK3
MARK4
 MELK
MNK1
 MNK2
 NIM1 kinase (MGC42105)
 NuaK1 (ARK5)
 PASK
 PhKg1
PhKg2
 Pim1 kinase
 Pim2 kinase
PKD1 (PKCµ)
PKD2
PKD3
 SIK
Ref.
➀
➀
➀
➀
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➁
➀ ➁
➁
➁
➁
➁
➁
➀ ➁
➁
➁
➁
➁
➁
➁
➀ ➁
➁
➁
➀ ➁
1572
2738
2739
2922
3024
2740
1582
2917
2882
1717
1930
2613
2741
2930
1466
1721
2684
1723
1966
2789
1967
2742
2614
2918
2792
2743
2744
2745
2622
2919
2920
2204
1729
1982
2921
kinase ASSAY LIST

January 2010
protein-serine/threonine kinases ❚ (cont'd)
Family/assay
protein-serine/threonine kinases / CaMK (cont'd)
smMLCK (MYLK)
➁
 STK33
➁
 TSSK1
➁
TSSK2 (STK22B)
protein-serine/threonine kinases / AGC
 Akt1/PKBa
➀ ➁
 Akt2/PKBb
➁
 Akt3/PKBg
➁
c  Akt1/2
CRIK
➁
GRK2 (ADRBK1)
➁
GRK3/BARK2 (ADRBK2)
➁
GRK5
MRCKa
➁
MSK1
➁
MSK2
➁
 NDR1 kinase
➁
 p70S6K
➁
 p70S6Kb
➁
 PDK1
➀ ➁
c  PDK1
 PKA
➀ ➁
PKCa
➁
 PKCb1
➁
 PKCb2
➀ ➁
PKCg
➁
PKCd
➁
PKCe
PKCz
➁
PKCh
➁
PKCq
➁
PKCi
PKG1a
➁
PKG1b
➁
PKG2
➁
PKN1
➁
 PKN2
➁
PRKX
➁
ROCK1
➀ ➁
 ROCK2
➁
RSK1
➁
 RSK2
➁
RSK3
➁
 SGK1
➀ ➁
other kinases ❚
Family/assay
other kinases
 AurA/Aur2 kinase
AurB/Aur1 kinase
AurC/Aur3 kinase
BubR1 kinase
 CDC7/ASK
 IKKa
 IKKb
IKKe (IKBKE)
 MOS kinase
MYT1 kinase
 NEK1
 NEK2
 NEK4
NEK6
NEK7
PEK (EIF2AK3)
PI3Kb
c
cellular assay

new assay
➁
➀ ➁
➁
➁
➁
➁
➀ ➁
➁
➁
➁
➁
new protocol
2026
3039
2791
2015
2923
2924
2925
G164
2628
2200
2627
2020
2629
1724
1983
2758
2883
2747
2926
G158
2927
0348
2888
2750
0350
0614
2044
2045
2046
2120
2121
2469
1730
1928
2615
2746
2681
1710
2884
2616
2928
2682
2929
Ref.
➀ ➁
➁
➁

Ref.

2885
1841
1842
2632
2764
2937
2938
2587
2765
2521
1931
2939
2950
1933
2588
2589
2203
Family/assay
protein-serine/threonine kinases / AGC (cont'd)
SGK2
 SGK3
➁
protein-serine/threonine kinases / CK1
CK1a
➀ ➁
CK1e
protein-serine/threonine kinases / STE
ASK1
➁
 COT kinase (MAP3K8)
➁
GCK (MAP4K2)
➁
HGK (MAP4K4)
➀ ➁
MEK1 (MAP2K1)
➁
c  MEK1
MEK5 (MAP2K5)
➁
 MEKK3 (MAP3K3)
➁
 MEKK4 (MAP3K4)
➁
MINK
➁
MKK4/JNK1
MKK4/JNK2
 MKK6
➀ ➁
MKK6/p38a (mutant active)
MST1 kinase (STK4)
➁
 MST2 kinase
➁
 MST3 kinase
➁
MST4 kinase
➀ ➁
NIK
➁
PAK1
➁
 PAK2
➀ ➁
PAK4
➀ ➁
TAOK2 (TAO1)
➀ ➁
protein-serine/threonine kinases / tkl
 ALK4
➁
 BMPR1A kinase (ALK3)
➁
B-raf kinase
➁
DLK1 (MAP3K12)
➁
IRAK1
➁
 IRAK4
➀ ➁
 LIMK1
➁
 LRRK2
 MLK1
➁
MLK2 (MAP3K10)
➁
 RAF-1 kinase
➁
RAF-1/MEK1
 RIPK2
➁
 TAK1-TAB1 (MAP3K7)
➁
Family/assay
other kinases (cont'd)
PKR (EIF2AK2)
PLK1
 PLK2
 PLK3
 PLK4
 TTK
 Wee1 kinase
 WNK2
WNK3
WNK4
assay converted from HTRF® to LANCEUltra®
1969
2790
1716
2630
2124
2003
2590
2021
0703
G159
2631
2760
2759
2113
1927
1953
2931
1679
2585
2761
2748
2024
2325
1934
2932
1935
2586
2115
2762
2016
2620
1968
2933
2934
3177
2935
2326
2936
1703
2763
3017
Ref.
➁
➀ ➁
➁
➁
➁
➁
➁
➁
➁
➁
atypical kinases ❚
Family/assay
atypical kinases
 mTOR kinase (FRAP1)
Ref.
2522
2049
2766
2793
2749
2767
2940
2768
2114
2633
Ref.
➀ ➁
2941
human ➀ ExpresS Diversity kinase profile ➁ Comprehensive kinase profile
3
kinase ASSAY LIST

January 2010
❚ testing conditions
❚ suggested testing
© Cerep - January 2010
Primary screening at 1-10 µM in duplicate (2 wells), followed-up for IC50 determination (8 concentrations in duplicate (16 wells)
when compound displays more than 50% inhibition of control value.
❚ sample size (including IC50 follow-up studies)
Assuming a molecular weight ≤ 500 g/mol and a testing concentration of 10 µM in duplicate (including a possible retest).
Screening
Weight (pre-weighed)
Screening + Follow up 1
Volume (100% DMSO)
Weight (pre-weighed) Volume (100% DMSO)
25 µL @ 10 mM
35 µL @ 10 mM
50 µL @ 10 mM
65 µL @ 10 mM
75 µL @ 10 mM
100 µL @ 10 mM
150 µL @ 10 mM
225 µL @ 10 mM
300 µL @ 10 mM
350 µL @ 10 mM
400 µL @ 10 mM
500 µL @ 10 mM
600 µL @ 10 mM
1 mg
1 mg
1 mg
1 mg
1 mg
1.5 mg
2 mg
2.5 mg
3 mg
4 mg
4 mg
5 mg
inquire
Individual catalog assays
1 to 3 assays
4 to 5 assays
6 to 10 assays
11 to 15 assays
16 to 20 assays
21 to 40 assays
41 to 50 assays
51 to 70 assays
71 to 100 assays
101 to 135 assays
136 to 150 assays
151 to 200 assays
201 to 250 assays
1
1 mg
1 mg
1 mg
1 mg
1 mg
1 mg
1 mg
1.5 mg
2 mg
2 mg
2.5 mg
3 mg
3.5 mg
60 µL @ 10 mM
70 µL @ 10 mM
100 µL @ 10 mM
110 µL @ 10 mM
125 µL @ 10 mM
250 µL @ 10 mM
275 µL @ 10 mM
400 µL @ 10 mM
550 µL @ 10 mM
650 µL @ 10 mM
750 µL @ 10 mM
1000 µL @ 10 mM
inquire
Assuming ~10% of test in IC50. Usually, for 1 IC50: 30 µL @ 10 mM and + 25 µL @ 10 mM by additional IC50.
❚ requested compound information
To reduce the registration time and ensure that all the appropriate information is available to start the study in a shortest possible
timeframe, please use Cerep compound submission form 1 or MS Excel file 2, and provide the following compound information:
 Name (compound ID) / Batch # / Molecular weight 3 / Formula weight
4/ Stock concentration / Stock solvent / Quantity /
Unit 5 / Form / Storage conditions / Solubility, as well as Plate ID / plate position for compounds delivered in plates, Comments 6,
and Quotation number.
NOTE: Impurity and colored compounds might affect the results (compound color information is mentioned in the study report).
Cerep compound submission
form will be emailed to you with
your quotation. A copy can be
requested from sales@cerep.
com, or downloaded from Cerep
website: www.cerep.com/
Catalog Online
2 Systematically required for studies
of 10 compounds or more.
3 Molecular weight (MW) of free
acid or base form.
4 Formula weight (FW) including
salt form and/or hydrate form if
applicable.
5 mg?, mL?
6 e.g. useful information such as
sensitivity to light, stability or
hygroscopicity issues.
1
 General remarks:
- If compound(s) are supplied as a stock solution in plate(s) (preferred format for any submission of 10 or more compounds),
please leave columns 1 and 12 empty in a 96W plate. The 384W plate format is also acceptable with columns 1, 2, 23
and 24 empty. For any other plate format, please inquire.
- If compound(s) are not soluble in 100% DMSO, please provide any useful information concerning the solubility of the compound.
The following solvents are compatible with most of our assays: DMSO (Cerep standard), H2O, Methanol, Tris/HCl 10 mM pH 7.4.
- Organic solvents such as acetone, chloroform, ether, acetonitrile, tetrahydrofuran and trifluoroacetic acid are not recommended
as they will significantly affect the results from many in vitro assays, even at very low concentrations.
Warning: Cerep will apply the standard solubilization process when compounds are received at the testing site, unless special
instructions are provided with the compounds.
Customized handling procedure of compounds can be accommodated, please inquire for pricing conditions.
❚ protocol
A typical protocol includes a minimum of 6-control wells (background, and maximal signal with and without vehicle) of the relevant
reference compound. The reference compound for each assay is listed in each assay description. The historical average IC50 value
is also shown in each assay description.
 Any of our assay protocols can be customized: Please inquire
❚ deliverables
Percent inhibition (mean of replicates), individual values as percent of control, IC50 value (calculated from a minimum 5 concentration testing), Hill coefficient (nH), and plotted IC50 curves.
❚ data turnaround
Complete data set is typically available within 3 weeks, after receipt of the compounds at the testing site (providing that we receive
all available information to initiate the study).
Secure, password-protected data can be viewed on line as soon as they are produced, after scientific approval and QC-ed by
an experienced technician.
FRANCE
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tel. +33 (0)5 49 89 30 00
(Headquarters)
155 boulevard Haussmann
75008 PARIS
tel. +33 (0)1 45 64 44 60
USA
15318 N.E. 95th Street
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tel. +1 (425) 895 8666
japan
Namiki Shoji Co., Ltd.
Kenseishinjuku Bldg. 5-5-3
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TOKYO, 160-0022
tel. +81 (0)3 3354 4026
fax +81 (0)3 3352 2196
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Zhangjiang Hich-Tech Park
Shanghai
tel. +86 18702160370
sales@cerep.com
www.cerep.com