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 爱游戏平台注册登录 :

• 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
• Kinase buffer suitable for you kinase (we do not recommend phosphate-based buffers)
• Tagged substrate
• Anti-phospho antibody
• ATP
• EDTA to stop kinase reaction

Instrumentation/Equipment:

• A TRF-capable plate reader

Top

Products and catalog numbers

View a listing of relevant reagents (with catalog numbers) for LANCE assays.

Top

Application notes and references for LANCE indirect kinase assays

Application note for an indirect-format LANCE kinase assay

1. 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
2. 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

Top