Ankyron PDK1_HUMAN Clone CD30228

HeLa were fixed in formaldehyde (1%), then stained using 1.4 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). Inset: negative control of anti-V5 APC antibody alone.

Immunohistochemical analysis of paraffin-embedded human liver using 10 μg/ml V5-tagged Ankyron and anti-V5 HRP (1 in 500), detecting with DAB (brown) and counterstaining nuclei with haematoxylin (blue-purple). Heat-mediated antigen retrieval with Leica Tris-EDTA buffer (pH9), 10 min.

Colorimetric western blot analysis of purified target protein (1 µg) using 1 µg/ml V5-tagged Ankyron preconjugated with anti-V5-HRP (1 in 500).

Binding assay determination of the signal to noise ratio (SNR) of V5-labeled Ankyrons® CD30228 binding to 0.5 picomoles of immobilised PDK1_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.

Affinity measurement of V5-labeled Ankyrons® CD30228 binding to PDK1_HUMAN, of value K_d = 31.9 nM, with 95% confidence bounds (CI95_U, CI95_L). Conjugation to ProImmune Anti-V5 HRP label reagent allows detection in Solution Equilibrium Titration immunoassay with a chemiluminescent substrate.