| Assay Method Information | |
| | Biochemical Assay |
| Description: | Compounds of the invention may be tested for in vitro activity in the following assay: A biotin labeled peptide is used as substrate (amino acid sequence: Biotin-Glu-Gly-Pro-Trp-Leu-Glu -Glu-Glu-Glu-Glu-Ala-Tyr-Gly-Trp-Met-Asp-Phe-NH2) (SEQ ID NO: 1). VEGFR3 cytoplasmic domain (amino acids 798-1298) was purchased as N-terminal GST-fusion protein (the enzyme). The 15ul assay reactions are run in Greiner brand white 384-well low volume plates. All reactions contained 10 mM HEPES pH 7.4, 10mM MgCl2, 0.01% (v/v) Tween-20, 50uM Na3VO4, 0.01% (w/v) albumin from chicken egg white, 1mM Dithiothreitol, 111 nM peptide substrate, 500uM ATP, and 3.8ng/reaction enzyme, with the enzyme being omitted from negative control reactions. Compounds were added in a volume of 100nl from dilution series prepared in DMSO, positive and negative control reactions receiving the same volume DMSO without compound. The plates were sealed with adhesive seals and incubated for 90 minutes at 30 degree C. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |