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AssayMetrics www.assaymetrics.com Bringing you fast, reliable fluorescence lifetime technologies
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PURETIME ® tyrosine kinase assays
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Key Features |
Key Benefits |
- Kinase and substrate additions
only
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- One step |
- No particles or antibodies |
- Cost effective |
- Simple fluorescence intensity or fluorescence lifetime |
- Use existing reader or upgrade to robust lifetime plate reading |
- Instant measurement of kinase activity |
- Kinase kinetics easily measured |

Fluorescence of PURETIME®17 is enhanced upon phosphorylation by the kinase enzyme, producing more intense emission and a longer fluorescence lifetime. The small fluorophor does not adversely affect the action of the kinase protein either binding ATP or phosphorylating the substrate. Fluorescence measurements can be made at any point during the kinase incubation, permitting time course experiments and direct determination of kinase reaction kinetics as well as end points.
Examples
Kinase activity assay


Lyn kinase enzyme titration performed in 384 well plate using EPEGIYGVLF-PURETIME17 substrate (500nM)
(intensity mode above, fluorescence lifetime mode below)
Kinase time course assay


Lck kinase time course performed in 384 well plate using EPEGIYGVLF-PURETIME17 substrate (500nM)
(intensity mode above, fluorescence lifetime mode below)
IC50 measurement assay

PP2 inhibition of Lyn kinase using EPEGIYGVLF-PURETIME17 substrate (1000nM)
(IC50 found to be 6nM and 40nM at 4 micM and 40micM ATP respectively)
Fluorescence lifetime discriminated measurements confirmed ATP analogue PP2 to exhibit competitive inhibition of Lyn kinase. Actual IC50 values are in good agreement with published values.
| Product code | N terminus | Sequence | C terminus | Amount |
| PT17-SRC-P | Ac- | EPEGIYGVLF | -
PURETIME
® 17 |
1mg |
| PT17-LCK-P | Ac- | KVEKIGEGTYGVV | -
PURETIME
® 17 |
1mg |
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