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- W4309751460 abstract "Abstract Polypeptide tags and biotin labelling technologies are widely used for protein analyses in biochemistry and cell biology. Although biotin labelling reacts with a lysine residue, many peptide tag epitopes have lysine residue(s). Here, we propose the GATS tag system without a lysine residue and with high sensitivity and low non-specific binding using a rabbit monoclonal antibody against Plasmodium falciparum glycosylphosphatidylinositol (GPI)- anchored micronemal antigen (PfGAMA). From 14 monoclonal clones, an Ra3 clone was selected as it recognized an epitope—TLSVGVQNTF—without a lysine residue; this antibody and epitope tag set was called the GATS tag system. Surface plasmon resonance analysis showed that the tag system had a high affinity of 8.71 × 10 -9 M. GATS tag indicated a very low background with remarkably high sensitivity and specificity in immunoblotting using the lysates of mammalian cells. It also showed a high sensitivity for immunoprecipitation and immunostaining of cultured human cells. The tag system was highly sensitive in both biotin labelling methods for proteins using NHS-Sulfo-biotin and BioID (proximity-dependent biotin identification) in the human cells, as opposed to a commercially available tag system having lysine residues, which showed reduced sensitivity. These results showed that the GATS tag system is suitable for methods such as BioID involving labelling lysine residues." @default.
- W4309751460 created "2022-11-29" @default.
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- W4309751460 date "2022-11-21" @default.
- W4309751460 modified "2023-10-17" @default.
- W4309751460 title "GATS Tag System for Protein Analysis with Biotin Labelling Methods" @default.
- W4309751460 doi "https://doi.org/10.21203/rs.3.rs-2254734/v1" @default.
- W4309751460 hasPublicationYear "2022" @default.
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