PURETIME ® tyrosine kinase assays
Key Features |
Key Benefits |
- Kinase and substrate additions
only
|
- 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 |
Kinase assay principle
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.
Products
| Product code | N terminus | Sequence | C terminus | Amount |
| PT17-SRC-P | Ac- | EPEGIYGVLF | -
PURETIME
® 17 |
1mg |
| PT17-LCK-P | Ac- | KVEKIGEGTYGVV | -
PURETIME
® 17 |
1mg |
Ordering
We ship direct from the UK all over the world. E-mail your request for a quotation or more information to sales@assaymetrics.com. Before placing an order with us, please ensure you have read our Terms and Conditions.