Overview
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 Revvity:
- 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™)
- Seal™-A adhesive plate seal
- 10X LANCE Detection buffer
Required reagents available from various suppliers:
- Kinase
- Kinase buffer suitable for your kinase (we do not recommend phosphate-based buffers)
- Tagged substrate
- Anti-phospho antibody
- ATP
- EDTA to kinase reaction
Instrumentation/Equipment:
- A TRF-capable plate reader
Products and catalog numbers
View a listing of relevant reagents (with catalog numbers) for LANCE assays.
Application notes and references for LANCE indirect kinase assays
For research use only. Not for use in diagnostic procedures.
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
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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