LANCE Indirect 4-Component Kinase Assays
The LANCE® indirect assay uses an unlabeled anti-phospho antibody with a Europium-labeled secondary antibody. This allows you to use your own unlabeled anti-phospho antibody with one of our catalog Eu-labeled secondary antibodies.
LANCE TR-FRET 4-component kinase assay.
What do I need to run this assay?
Required reagents available from PerkinElmer:
- LANCE Europium-labeled reagent to bind anti-phospho antibody (alternatively, you can Label your own LANCE reagent)
- Acceptor fluorophore-labeled reagent to bind substrate
- Microplate (we recommend our white OptiPlates™)
- TopSeal™-A adhesive plate seal
- 10X LANCE Detection buffer
Required reagents available from various suppliers:
- Kinase buffer suitable for you kinase (we do not recommend phosphate-based buffers)
- Tagged substrate
- Anti-phospho antibody
- EDTA to stop kinase reaction
- A TRF-capable plate reader
Products and catalog numbers
Application notes and references for LANCE indirect kinase assays
Application note for an indirect-format LANCE kinase assay
- Bader, B. et al. A cGMP-Dependent Protein Kinase Assay for High Throughput Screening Based on Time-Resolved Fluorescence Resonance Energy Transfer. J Biomol Screen 6, 255-264 (2001). Link
- Zhang, W., Shor, B. & Yu, K. Identification and Characterization of a Constitutively T-Loop Phosphorylated and Active Recombinant S6K1: Expression, Purification, and Enzymatic Studies in a High Capacity Non-Radioactive TR-FRET Lance assay. Protein Expres Purif 46, 414-420 (2006). Link