The HTRF KinEASE assay series is a semi-universal biochemical platform to study kinase activity. The STK1 kit uses a specific substrate compatible with various Serine/thronine kinases - cf. validation data provided on this page.
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  • Low enzyme consumption Low enzyme consumption
The HTRF KinEASE assay series is a semi-universal biochemical platform to study kinase activity. The STK1 kit uses a specific substrate compatible with various Serine/thronine kinases - cf. validation data provided on this page.
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Overview

The HTRF KinEASE kit series provides a simple biochemical approach to study kinase activity, screen for inhibitors and characterize them. Three STK and one TK kits, offer a semi-universal method for assessing phosphorylation on Serine/Threonine and Tyrosine residues, respectively. The HTRF KinEASE STK S1 assay has been validated on 56 Serine/Threonine kinases. The assay validation section of this page lists them and provides technical tips for setup. The kit series also include a Discovery kit to help determine which of three STK kits is appropriate when working with a non-validated enzyme.

Benefits

  • COMPATIBLE WITH UP TO MM ATP KMS
  • SEMI-UNIVERSAL METHOD FOR STKS
  • LOW ENZYME CONSUMPTION

Assay principle

Each HTRF KinEASE kit is based on the same assay format, and uses a universal biotinylated substrate, a phospho-specific Eu-cryptate-labeled monoclonal antibody, and SA-XL665. All three STK peptidic substrates are made of the same sequence for antibody detection, next to a variable sequence recognized by enzymes. The targeted residue for phosphorylation lies in between as part of both sequences for enzymatic activity (unphosphorylated) and antibody detection (phosphorylated). It has been shown that 3 different variable sequences could enable the study of most Serine/Threonine kinases.

Assay protocol

As shown here, all HTRF KinEASE assays work the same way and involve two steps:

Step 1: Kinase reaction The kinase is incubated in the presence or absence of compounds and the appropriate substrate. The supplemented enzymatic buffer is added in the kinase buffer for tyr kinases only. ATP is added to start the reaction.

Step 2: Detection Eu3+ cryptate and XL665 conjugates are added. The detection buffer contains EDTA to stop the enzymatic reaction.

HTRF KinEASE Assay principle
Click in the table below to view the list of enzymes, the kit whose use is recommended, and to download the technical datasheet including experimental setting and validation data. Non listed STKs can be assessed with the Discovery kit to find out which HTRF KinEASE STK kit may be used.
ABL1 (TK)ABL2 (TK)AKT1 (STK S3)AKT2 (STK S3)AKT3 (STK S3)
ALK (TK)AMPK a2/ß1/g1 (STK S1)AMPK a2/ß2/g1 (STK S1)Arg (TK)ARK5 (STK S3)
ASK1 (STK S3)AURKA (STK S2)AURKB (STK S2)Aurora A (STK S2)Aurora B (STK S2)
Aurora C (STK S3)AXL (TK)BLK (TK)BMX (TK)BRK (TK)
BrSK1 (STK S3)BrSK2 (STK S1)BTK (TK)c-Kit (TK)CAMK1 (STK S1)
CaMK1 a (STK S1)CaMK2 a (STK S1)CaMK2 ß (STK S1)CaMK2 d (STK S1)CaMK2 g (STK S1)
CAMK2B (STK S1)CAMK2D (STK S1)CAMK2G (STK S1)CaMK4 (STK S1)CDC42 BPA (STK S3)
CDC42 BPB (STK S3)CGK2 (STK S2)CHEK1 (STK S1)CHEK2 (STK S1)CHK1 (STK S1)
CHK2 (STK S1)CHUK (STK S3)CLK3 (STK S3)COT (STK S3)CSF1R (TK)
CSK (TK)DAPK1 (STK S1)DAPK2 (STK S1)DAPK3 (STK S1)DCAMKL2 (STK S1)
DDR2 (TK)DMPK (STK S3)DRAK1 (STK S1)DYRK2 (STK S3)EEF-2K (STK S3)
EGFR (TK)EPHA1 (TK)EPHA2 (TK)EPHA3 (TK)EPHA4 (TK)
EPHA5 (TK)EPHA7 (TK)EPHA8 (TK)EPHB1 (TK)EPHB2 (TK)
EPHB3 (TK)EPHB4 (TK)ERBB4 (TK)FAK (TK)FER (TK)
FES (TK)FGFR1 (TK)FGFR2 (TK)FGFR3 (TK)FGFR4 (TK)
FGR (TK)FLT1 (TK)FLT3 (TK)FLT4 (TK)FMS (TK)
Fps (TK)FRK (TK)FYN (TK)GPRK4 (STK S1)GRK4 (STK S1)
GRK5 (STK S3)GRK6 (STK S1)GSG2 (STK S1)GSK3 a (STK S3)Haspin (STK S1)
HCK (TK)HER4 (TK)HIPK2 (STK S3)HIPK3 (STK S3)IGF1R (TK)
IKBKB (STK S3)IKK a (STK S3)IKK ß (STK S3)INSR (TK)INSRR (TK)
Insulin R (TK)IRAK4 (STK S1)IRR (TK)ITK (TK)JAK1 (TK)
JAK2 (TK)JAK3 (TK)KDR (TK)KIT (TK)LATS1 (STK S1)
LCK (TK)LKB1 (STK S1)LOK (STK S3)Lyn (TK)LYNa (TK)
MAP3K8 (STK S3)MAPKAP-K1 a (STK S1)MAPKAP-K1 ß (STK S1)MAPKAP-K2 (STK S1)MAPKAP-K3 (STK S1)
MAPKAP-K5 (STK S1)MARK1 (STK S1)MARK2 (STK S2)MELK (STK S1)MER (TK)
MERTK (TK)MET (TK)MINK1 (STK S3)MKNK1 (STK S1)MKNK2 (STK S1)
MLCK (STK S1)MNK1 (STK S1)MNK2 (STK S1)MRCK a (STK S3)MRCK ß (STK S3)
MSK1 (STK S3)MSK2 (STK S3)MSSK1 (STK S3)MST1 (STK S3)MST1R (TK)
MST2 (STK S3)MSTIR (TK)MUSK (TK)MYLK (STK S1)NEK11 (STK S2)
NEK2 (STK S3)NEK3 (STK S3)NEK6 (STK S3)NEK7 (STK S3)NLK (STK S3)
NTRK1 (TK)NTRK2 (TK)NTRK3 (TK)p70S6K (STK S3)PAK2 (STK S2)
PAK3 (STK S2)PAK4 (STK S2)PAK5 (STK S2)PAK6 (STK S2)PAK7 (STK S2)
PAR-1B a (STK S2)PASK (STK S1)PDGFR a (TK)PDGFR ß (TK)PDGFRA (TK)
PDGFRB (TK)PHK g1 (STK S1)PHK g2 (STK S1)PIM1 (STK S3)PIM2 (STK S3)
PKAC a (STK S2)PKAC g (STK S3)PKB a (STK S3)PKB ß (STK S3)PKB g (STK S3)
PKC a (STK S1)PKC ß1 (STK S1)PKC ß2 (STK S1)PKC d (STK S1)PKC epsilon (STK S1)
PKC µ (STK S1)PKC theta (STK S1)PKC eta (STK S1)PKC gamma (STK S1)PKC iota (STK S1)
PKC zeta (STK S1)PKD2 (STK S1)PKG1 a (STK S2)PKG1 ß (STK S2)PKG2 (STK S1)
PKN2 (STK S1)PLK1 (STK S1)PLK3 (STK S3)PLK4 (STK S2)PRAK (STK S1)
PRK2 (STK S1)PRKA A2/B1/G1 (STK S1)PRKACA (STK S2)PRKACG (STK S3)PRKCA (STK S1)
PRKCB1 (STK S1)PRKCB2 (STK S1)PRKCD (STK S1)PRKCE (STK S1)PRKCG (STK S1)
PRKCH (STK S1)PRKCI (STK S1)PRKCQ (STK S1)PRKCZ (STK S1)PRKD2 (STK S1)
PRKG1 (STK S2)PRKG2 (STK S1)PRKX (STK S2)PTK2 (TK)PTK2B (TK)
PTK5 (TK)PTK6 (TK)PYK2 (TK)RET (TK)ROCK1 (STK S2)
ROCK2 (STK S2)ROK a (STK S2)RON (TK)ROS (TK)ROS1 (TK)
RPS6KA1 (STK S1)RPS6KA2 (STK S1)RPS6KA3 (STK S1)RPS6KA4 (STK S3)RPS6KA5 (STK S3)
RPS6KA6 (STK S2)RPS6KB1 (STK S3)Rse (TK)RSK1 (STK S1)RSK2 (STK S1)
RSK3 (STK S1)RSK4 (STK S2)SAD1 (STK S3)SGK1 (STK S3)SGK2 (STK S3)
SGK3 (STK S3)SGKL (STK S3)SIK (STK S1)smMLCK (STK S1)Snk (STK S3)
SRC (TK)STK22B (STK S1)STK22D (STK S1)STK23 (STK S3)STK3 (STK S3)
STK4 (STK S3)STK6 (STK S2)SYK (TK)TBK1 (STK S1)TEK (TK)
TIE2 (TK)TRKA (TK)TRKB (TK)TRKC (TK)TSSK1 (STK S1)
TSSK2 (STK S1)TYK2 (TK)TYR03 (TK)TYRO3 (TK)VEGFR1 (TK)
VEGFR2 (TK)VEGFR3 (TK)WNK2 (STK S3)WNK3 (STK S3)YES1 (TK)
ZAP70 (TK)ZIPK (STK S1)

Validated kinases

Over 190 Serine/Threonine and Tyrosine kinases have already been validated with the HTRF KinEASE kits. Out of those, 56 were found to be compatible with the S1 kit. The case of SIK Serine/Threonine kinase is illustrated below with the enzyme titration and time course (on the left), and the inhibition by staurosporine (on the right).
SIK kinase titration and time course
SIK kinase inhibition by staurosporine
CaMK1-alpha, CAMK1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

CaMK2-beta, CAMK2B validated with HTRF KinEASE

Enzyme validation - Technical Notes

CaMK2-gamma, CAMK2G validated with HTRF KinEASE

Enzyme validation - Technical Notes

CaMK2-delta, CAMK2D validated with HTRF KinEASE

Enzyme validation - Technical Notes

CHK1, CHEK1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

CHK2, CHEK2 validated with HTRF KinEASE

Enzyme validation - Technical Notes

DAPK1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

DAPK3, ZIPK validated with HTRF KinEASE

Enzyme validation - Technical Notes

DCAMKL2 validated with HTRF KinEASE

Enzyme validation - Technical Notes

GPRK4, GRK4 validated with HTRF KinEASE

Enzyme validation - Technical Notes

GRK6 validated with HTRF KinEASE

Enzyme validation - Technical Notes

Haspin, CSG2 validated with HTRF KinEASE

Enzyme validation - Technical Notes

IRAK4 validated with HTRF KinEASE

Enzyme validation - Technical Notes

LATS1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

LCK validated with HTRF KinEASE

Enzyme validation - Technical Notes

MAPKAP-K2 validated with HTRF KinEASE

Enzyme validation - Technical Notes

MAPKAP-K3 validated with HTRF KinEASE

Enzyme validation - Technical Notes

MAPKAP-K5, PRAK validated with HTRF KinEASE

Enzyme validation - Technical Notes

MARK1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

MELK validated with HTRF KinEASE

Enzyme validation - Technical Notes

MNK1, MKNK1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

MNK2, MKNK2 validated with HTRF KinEASE

Enzyme validation - Technical Notes

PASK validated with HTRF KinEASE

Enzyme validation - Technical Notes

PHK-gamma1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

PHK-gamma2 validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKC-alpha, PRKCA validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKC-beta1, PRKCB1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKC-beta2, PRKCB2 validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKC-gamma, PRKCG validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKC-delta, PRKCD validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKC-epsilon, PRKCE validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKC-zeta, PRKCZ validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKC-eta, PRKCH validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKC-theta, PRKCQ validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKC-iota, PRKC1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

PKD2, PRK2 validated with HTRF KinEASE

Enzyme validation - Technical Notes

PLK1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

PRK2, PKN2 validated with HTRF KinEASE

Enzyme validation - Technical Notes

RSK3, RPS6KA2 validated with HTRF KinEASE

Enzyme validation - Technical Notes

SIK validated with HTRF KinEASE

Enzyme validation - Technical Notes

smMLCK, MYLK validated with HTRF KinEASE

Enzyme validation - Technical Notes

TSSK2, STK22B validated with HTRF KinEASE

Enzyme validation - Technical Notes

TSSK1, STK22D validated with HTRF KinEASE

Enzyme validation - Technical Notes

TBK1 validated with HTRF KinEASE

Enzyme validation - Technical Notes

HTRF KinEASE

Universal platform to address Ser/Thr & Tyr kinases - Application Notes

Open R&D: Sanofi Access Platform

In collaboration with Sanofi - Scientific Presentations

Identification of natural ligands for AMP-activated protein kinase with HTRF

In collaboration with Nestlé Institute of Health Sciences - Posters

KinEASE, a universal expanded platform to adress Ser/Thr & Tyr kinases

Measuring kinase activity under any conditions - Application Notes

Exploration is easier with a trusted partner

HTRF KinEASE Tyr and Ser/Thr kits - Flyers

Cisbio KinEASE

Validated kinases and assay conditions - Flyers

Product Insert KinEASE STK S1 Kit / 62ST1PEB-62ST1PEC-62ST1PEJ

62ST1PEB-62ST1PEC-62ST1PEJ - Product Insert

Cisbio Product Catalog 2019

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

The essential white paper to expand your knowledge on kinases

Learn about cutting edge kinase research in this free white paper - Guides

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

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