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Tel: 1-631-448-8149
Fax: 1-631-938-8127

Cell-based Kinase Assays

Protein kinase represents an important family of target in drug development. The success of recently approved kinase inhibitors has proved the value of targeting kinases in cancer treatment. Despite the great progress in developing reagents and assays to measure kinase activity with purified proteins, questions remain about whether these methods can deliver sensitive, robust and accurate results. Unlike in vitro assays, cell-based methods provide direct information of how drug molecules truly interact with kinases in the presence of myriad known or unknown cellular factors, resulting in more reliable and relevant data.

For years, Profacgen has gleaned valuable experience from studies of numerous biological targets and systems. To meet the growing demand in kinase study, we are now offering a novel platform to evaluate cell-based activity of drug candidates with a focus on a comprehensive set of critical parameters. At Profacgen, cell-based kinase assays are offered in two ways:

1) Activity measurement by phosphorylation level. We use transient expression technique to introduce kinases into human cells, and quantify their phosphorylation levels by antibody-antigen reactions. This strategy, originally built upon human embryonic kidney cells, allows for quick potency evaluation as well as mechanistic studies. Reduced phosphorylation level can be easily detected by specific antibody and provides a strong sign for kinase inhibition.

2) Activity measurement by cell survival. Pioneered by Daley and Baltimore,1 this method intends to express kinases of interest in IL3-dependent Ba/F3 cells. Upon removal of IL3, cell proliferation and survival is dependent on the activity of the introduced kinase. Change in cell viability after drug administration is associated with inhibitory effect and indicates loss in kinase activity.

We proudly offer a large selection of kinase targets in cell-based assays:

Tyrosine Kinases
ABL (BCR-ABL) EphA1 FGFR3 HER2 (ERBB2) PDGFRβ
ALK EphA3 FGFR3 [K650M] HER3 (ERBB3) RET
ARG (ABL2) EphA4 FGFR3/BAIAP2L1 IGF1R RET [V804M]
AXL EphA5 FGFR4 INSR RON (MST1R)
BLK EphB1 FGFR4 [V550E] JAK1 ROR1
BMX EphB2 FGR JAK2 ROS (ROS1)
BTK EphB4 FLT1 (VEGFR1) JAK3 RYK
CCK4 (PTK7) FAK FLT3 KDR (VEGFR2) SRC
DDR2 FGFR1 FLT3-ITD KIT SYK
EGFR FGFR1 [V561M] FLT3-ITD [D835V] KIT [D816V] TIE1
EGFR [D746-750] FGFR2 FLT3-ITD [D835Y] KIT [K642E] TIE2
EGFR
[D746-750+T790M]
FGFR2 [K660E] FLT3-ITD [F691L] KIT [N822H] TRKA (NTRK1)
EGFR [L858R] FGFR2 [K660N] FLT3-ITD [Y842C] KIT [T670I] TRKB (NTRK2)
EGFR
[L858R+T790M]
FGFR2 [N550K] FLT3-ITD [Y842H] KIT [V654A] TRKC (NTRK3)
EGFR [L858R+C797S] FGFR2 [V565I] FLT4 (VEGFR3) LCK TYK2
EGFR
[L858R+T790M+C797S]
FGFR2/AFF3 FMS (CSF1R) LYN TYRO3
EGFR [L861Q] FGFR2/BICC1 FRK MER (MERTK) ZAP70
Serine and Threonine Kinases
AKT1 DCLK2 DYRK1A MST1 PDK1
PIM1 PIM2 PIM3    

For more information about our cell-based kinase assays, please contact us for details. Our tech representatives are available to help you 24 hours a day, Monday through Friday.

Reference:

1. Daley, Q.; Baltimore, D. Transformation of an interleukin 3-dependent hematopoietic cell line by the chronic myelogenous leukemia-specific P210 bcr/abl protein. Proceedings of the National Academy of Sciences 1988, 85 (23): 9312-9316.

CONTACT US
Email: info@profacgen.com
Tel: 1-631-448-8149
Fax: 1-631-938-8127
Address: 45-1 Ramsey Road, Shirley, NY 11967, USA