Alphascreen Assay: The AlphaScreen assay was performed according to the manufacturer's protocol (Perkin Elmer, Benelux). Reactions were performed in 2...

ID: ALA3706055

Type: Binding

Description: Alphascreen Assay: The AlphaScreen assay was performed according to the manufacturer's protocol (Perkin Elmer, Benelux). Reactions were performed in 25 ul final volume in 384-well Optiwell microtiter plates (Perkin Elmer). The reaction buffer contained 25 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM MgCl2, 0.01% (v/v) Tween-20 and 0.1% (w/v) bovine serum albumin. His6-tagged integrase (300 nM final concentration) was incubated with the compounds for 30 min at 4C. The compounds were added at varying concentrations spanning a wide range from 0.1 up to 100 uM. Afterwards 100 nM flag-LEDGF/p75 was added and incubation was prolonged for an additional hour at 4C. Subsequently 5 ul of Ni-chelate-coated acceptor beads and 5 ul anti-flag donor beads were added to a final concentration of 20 ug/ml of both beads. Proteins and beads were incubated for 1 h at 30C. in order to allow association to occur.

Format: BAO_0000357

Target:  Gag-Pol polyprotein(ALA3638326)

Document:  ALA3638570

BindingDB Source:  6398