iBody anti-polyHis 800 Targeting Variably Long Polyhistidine Fusion Tags
designed for near-infrared fluorescent detection
Anti-polyHis 800 iBody is able to specifically target and bind polyhistidne fusion tags. The so called His-tag, commonly engineered to recombinant proteins, is the most frequently used affinity tag utilized for purification, detection and characterization of such molecules. The ligand used for specific targeting is the tris-trinitriloacetic acid (tris-NTA) which is able to bind the polyhistidine sequence through the formation of a stable complex via a metal cation mediator. iBody anti-polyHis 800 is also accessorized with Cyanine fluorophore, which serves as a tool for facile and fast detection at near IR region. Furthermore, biotin is conjugated to the polymer carrier. It assists in immobilization of the protein of interest and potentially in additional detection with chemiluminesce.
anti-polyHis iBodies serve as a replacement for monoclonal anti-His antibodies in a variety of biochemical applications such as western blotting (WB), ELISA, flow cytometry, confocal fluorescent microscopy, protein immobilization on resin etc.
Western Blotting with anti-polyHis 800:
Fig.1: Detection of 10xHis and 6xHis-tagged proteins on western blot. Comparison of detection with commercially available mAbs.
Targeted biomolecule specification:
Protein containing polyhistidine sequence (6×His, 10×His - the most widely used purification tags) fused to the N- or C-terminus of a protein of interest.
Targeting ligand: tris-NTA (tris-nitrilotriacetic acid) charged with cobalt cations
Affinity anchor: biotin
Mr ~ 87 kDa; Cyanine 7.5 fluorophore (2 wt%; 2 molecules), biotin content (8 wt%; 24 molecules), tris-NTA ligand (16 wt%; 11 molecules).
iBody KD = 4 nM (10xHis-protein as a model system; measured with surface plasmon resonance)
The iBody is stable at room temperature. For long term storage, place the solution to -20°C.
Angew Chem Int Edit 2016, 55, 2356-2360