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LANCE Indirect 4-Component Kinase Assays

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_4_component_ASK.jpg
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
  • 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

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Products and catalog numbers


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

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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

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