KinEASE
Non-Radioactive Kinase Assays
KinEASE is a robust fluorescence polarization kinase assay optimized for use with 80 different kinases. KinEASE is optimized for low volume assays for which there are no practical limits on ATP concentration, making it ideal for use in kinase inhibitor identification and potency studies.
PDF: Fluorescence Polarization Kinase Activity Assays
| Abl (H) | FGFR3 (H) | PDGFRβ (H) | Ret (H) |
| Abl (M) | Flt1 (H) | Pim-1 (H) | ROCK-II (H) |
| Abl (T3151) (H) | Flt3 (H) | PKA (H) | Ros (H) |
| AMPK (R) | Fms (H) | PKBα (H) | Rse (H) |
| Arg (H) | Fyn (H) | PKBβ (H) | Rsk1 (H) |
| Arg (M) | IGF-1R (H) | PKBγ (H) | Rsk1 (R) |
| Aurora A (H) | IKKα (H) | PKCα (H) | Rsk2 (H) |
| Blk (M) | IKKβ (H) | PKCβ I (H) | Rsk3 (H) |
| Bmx (H) | IR (H) | PKCβ II (H) | SAPK2β (H) |
| BTK (H) | Lck (H) | PKCδ (H) | SAPK2β (H) |
| CaMKII (R) | Lyn (H) | PKCε (H) | SAPK3 (H) |
| CaMKIV (H) | Lyn (M) | PKCγ (H) | SAPK4 (H) |
| Chk1 (H) | MAPK1 (H) | PKCη (H) | SGK1 (H) |
| Chk2 (H) | MAPK2 (H) | PKCι (H) | Src (H) |
| CSK (H) | MAPKAP-K2 (H) | PKCμ (H) | Syk (H) |
| EFGR (H) | Met (H) | PKCθ (H) | Tie2 (H) |
| EphA2 (H) | MSK1 (H) | PKξ (H) | TrkA (H) |
| EphB2 (H) | p7056K (H) | PKD2 (H) | TrkB (H) |
| EphB4 (H) | PAK2 (H) | PRAK (H) | Yes (H) |
| Fes (H) | PDGFRα (H) | PRK2 (H) | ZAP-70 (H) |
KinEASE is designed to reduce the compound interference through the use of Upstate's proprietary FarRed fluorescent tracer. KinEASE also offers the significant advantage of ratiometric measurement, resulting in high quality, reproducible data. The phospho-specific monoclonal antibodies used in this assay, including our best selling 4G10®, have already been optimized, saving you time and costly reagents.
Each KinEASE reagent pack can generate up to 800 data points and is available for single kinase targets as well as with a combination of reagents to allow for screening of multiple serine/threonine and tyrosine kinases. All KinEASE kit components are available in bulk quantities as well as sold individually to allow further customization of your research options.
Principle Of FP Homogeneous Assay Format
Top panel:
Kinase is added to wells containing 0.5 µlof compound in 100% DMSO. The enzyme reaction is then started by addition of ATP/Mg and incubated at room temperature for 40 minutes.Middle panel:
The kinase reaction is stopped by the addition of EDTA plus monoclonal antibody and a fluorescent phospho-tracer, i.e., the detection module.Bottom panel:
Detection of the polarized signal. Kinase activity yields a product that competes with the fluorescent-phospho- tracer for binding to the anti-phospho-mAb, resulting in a low polarization signal. If the kinase is inhibited there is no competition and thus a high polarization signal is observed.
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