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Histidine (His) tags are widely used for affinity purification of recombinant proteins in a nickel column.
Chemically synthesized ligands identified by this screening were immobilized onto a chromatographic support and used for affinity purification of factor VIII from a complex feedstream.
Here we show that robust and small protein ligands can be used for affinity capture of recombinant proteins from crude cell lysates.
The pBpa group was then used for affinity capture of the mutated antibody by β-cyclodextrin (β-CD), which provided the hydrogen atoms to be abstracted in the subsequent photocoupling process upon irradiation at 365 nm.
Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification.
They cannot only be used for affinity purification but also downstream for detection and assay of an arbitrary fused recombinant protein without the need for any prior knowledge of its biochemical properties.
When immobilized on a matrix, the antibody can be used for affinity purification of recombinant proteins carrying a very short tag of just three histidine residues, suitable for crystallization.
The final bleed was used for affinity purification.
A 20 ml bed volume CaM-sepharose column was used for affinity purification.
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The Nub peptide without the carrier protein was coupled to cyanogen bromide-activated sepharose 4B according the manufacturer's protocol (Sigma-Aldrich, St Louis, MI, USA), and used for affinity-purification of the antisera.
In addition, midpoints or medians were used for affinities expressed as ranges.
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