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Ankyron RCAS1_HUMAN Clone BA40127
Ankyrons® Testing Data
HeLa cells were stained using 10 µg V5-tagged Ankyron and anti-V5 antibody conjugated with APC (red) in Permeabilization Staining Buffer (RPMI 1640 + 10 % human serum + 0.5 % saponin (from quillaja bark, Sigma-Aldrich, w/v in PBS)), counterstaining nuclei with DAPI (blue), (BioLegend) and mounting the slide with VECTASHIELD® PLUS Antifade Mounting Medium. Image captured by epifluorescence microscopy using an Echo Revolve microscope.
HUVEC were fixed in formaldehyde (4%), 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) and F-actin with Phalloidin. Inset: negative control of anti-V5 APC antibody alone and F-actin staining.
HUVEC fixed in methanol, then stained using 1.4 µg V5-tagged Ankyron and anti-V5 APC (red) in Staining Buffer (RPMI 1640 + 10 % human serum), counterstaining nuclei with DAPI (blue). Inset: negative control of anti-V5 APC alone.
HeLa cells were fixed in formaldehyde (1%), then stained using 1.4 µg V5-tagged Ankyron and anti-V5 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 alone.
Flow cytometric analysis of human HeLa cells following permeabilization and staining with pre-conjugated Ankyron (0.4 µg) and anti-V5 APC (red). Negative control, anti-V5 APC only (grey).
Flow cytometric analysis of mouse J774A.1 following permeabilization and staining with pre-conjugated Ankyron (0.4 µg) and anti-V5 APC (red). Negative control, anti-V5 APC only (grey).
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® BA40127 binding to 0.5 picomoles of immobilised RCAS1_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.