HUVEC were cultured for 20 h in either the absence or presence of TNF-α, 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.
HUVEC 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.
Immunohistochemical analysis of paraffin-embedded human tonsil 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 EDTA buffer (pH9), 40 min.
Flow cytometric analysis of HUVEC following permeabilisation with Cytofix/Cytoperm and staining with pre-conjugated Ankyron (0.4 µg) and anti-V5 APC (red). Negative control, anti-V5 APC only (grey).
Binding assay determination of the signal to noise ratio (SNR) of V5-labeled Ankyrons® AB30108 binding to 0.5 picomoles of immobilised CCL2_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.
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