Ankyron TF65_HUMAN Clone AW40914

HUVEC were cultured overnight in either the absence (control, unstimulated) or presence of TNFα, to track translocation of target protein from cytoplasm to nucleus in response to TNFα stimulation. Cells were permeabilised with 0.1% Triton X-100 and then stained using 2.5 µg V5-tagged Ankyron and anti-V5 antibody conjugated with APC (red) in Permeabilisation Staining Buffer (RPMI 1640 + 10 % human serum + 0.5 % saponin), counterstaining nuclei with DAPI (blue) and F-actin with PhalloidinImage captured by epifluorescence microscopy. Insets: negative control of anti-V5 APC antibody alone, with DAPI and F-actin counterstaining.

Binding assay determination of the signal to noise ratio (SNR) of V5-labeled Ankyrons® AW40914 binding to 0.5 picomoles of immobilised TF65_HUMAN, 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.