Ankyron NEF_HV1BR Clone DV40751

HEK293T cells were transfected to express HIV-1 Nef from a Nef IRES GFP-encoding plasmid then harvest by trypsinisation, 24h later. Cells were washed, fixed in 4% PFA in PBS for 10 min at RT (approx. 50k-100k cells per staining condition), washed again and then permeabilised with ice cold 90 % methanol on ice for 15 min, prior to staining with Ankyron preconjugated with anti-V5 APC labeled Fluorotag. During flow cytometric analysis, live, single cells were gated for Nef GFP+ and Ankyron APC+ populations. Data are courtesy of Dylan Postmus, London School of Tropical Medicine.

Binding assay determination of the signal to noise ratio (SNR) of V5-labeled Ankyrons® DV40751 binding to 0.5 picomoles of immobilised NEF_HV1BR, over a background (BG) of non-cognate target protein. The Ankyron was pre-conjugated with the ProImmune Anti-V5 HRP label reagent before detection with a chemiluminescence substrate.