VEGF Binding to the VEGFR2 Assessed in Real-Time in Living Cells with NanoBRET™ Assay

Growth factors stimulate receptor tyrosine kinases (RTK) which results in regulation of cell proliferation, growth, differentiation, and cell death. Therefore, RTKs are an attractive anti-cancer target as aberrant RTK-signaling promotes cancer. Expansion of pharmaceutical RTK-inhibitors needs methods which characterize receptor-ligand interaction and identify the inhibitors of the interaction.

Using an innovative technique, the vascular endothelial growth factor (VEGF) was tagged at a single locus with the fluorophore TMR. The labeled ligand was used in conjugation with HEK293 cells exhibiting the RTK VEGF-receptor 2 (VEGFR2) attached to the NanoLuc® luciferase. Upon interaction between receptor and ligand, bioluminescence resonance energy transfer (BRET) between luciferase and fluorophore occurs.

The PHERAstar microplate reader obtained the signal of luciferase and fluorophore in parallel. In this manner, not only the pKi of a VEGF-inhibitor was measured, but also the time-course of inhibition and binding was observed.

Read the full article including the methods, results and discussion.

NanoBRET assay quantitatively evaluates VEGF binding to the VEGFR2 in real time in living cells

About BMG Labtech

BMG LABTECH has been committed to producing microplate readers for more than twenty years. By focusing on the needs of the scientific community, the company’s innovative microplate readers have earned the company the reputation of being a technology leader in the field.

BMG LABTECH has developed a wide range of dedicated and multi-mode microplate readers for life sciences applications and high-throughput screening.

All BMG LABTECH microplate readers are "Made in Germany" and are conceived, developed, assembled, and tested entirely at our headquarters in Germany.

Since our establishment in Offenburg, Germany in 1989, BMG LABTECH has expanded to offer a worldwide sales and support network with offices in the USA, UK, Australia, Japan and France. Our subsidiaries, regional offices and distributors are committed to bringing you innovative microplate reader technology with the quality and reliability you expect from a German company.

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Last updated: Jul 31, 2018 at 3:25 AM

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