Single-domain antibody n3113v labeled by DyLight 800 antibody Labeling Kit (ThermoFisher) at 12 mg/kg was administrated through inhalation or intraperitoneal injection. The single-domain antibody was administrated through intratracheal route using microsprayer aerosolizer (YUYANBIO). Briefly, before performing trachea cannula, mice mouth was opened up by the laryngoscope to visualize the porch of the trachea. The tip syringe of high-pressure microsprayer was gently fed into the main trachea, and the aerosols were delivered quickly. Mice were imaged at different time-points (0.5 h, 1 h, 1.5 h, 2 h, 2.5 h, 4 h, 6 h) and the fluorescence radiance was captured by IVIS Lumina K Series III (PerkinElmer) at Ex/Em 780 nm/845 nm. At 4 hours and 6 hours post antibody administration, mice were sacrificed and organs were collected for fluorescence imaging.
Dylight 800 antibody labeling kit
Manufactured by Thermo Fisher Scientific
The DyLight 800 antibody Labeling Kit is a tool designed for the fluorescent labeling of antibodies. It allows for the conjugation of DyLight 800 dye to antibodies, enabling their detection and visualization in various analytical techniques.
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2 protocols using dylight 800 antibody labeling kit
1
In Vivo Biodistribution of Labelled Antibody
2
Purification of PCSK9 and LDLR-ECD
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FLAG epitope-tagged recombinant human WT PCSK9 along with A44P, R46L, R496W, and Δ31–52 variants were produced in stably transfected HEK293S cells and purified from conditioned culture medium as described previously (18 (link)). Fluorescently labeled WT PCSK9 was prepared using the DyLight800 antibody-labeling kit (Thermo Fisher Scientific) as per the manufacturer's instructions followed by gel filtration chromatography on a Superdex 200 10/300 GL column (GE Healthcare) to remove unbound dye. The extracellular domain (ECD) of LDLR used for PCSK9 binding assays was partially purified from conditioned medium of HEK293S cells cultured as described (18 (link)) stably transfected with a plasmid encoding amino acids 1–692 of human LDLR containing a C-terminal His6 tag (a kind gift from R. Milne, University of Ottawa). LDLR-ECD was enriched by affinity chromatography using TALON superflow affinity resin (Clontech) followed by gel filtration chromatography on a Superdex 200 10/300 GL column (GE Healthcare).
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