2014
DOI: 10.1016/b978-0-12-420070-8.00007-6
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Labeling of a Protein with Fluorophores Using Maleimide Derivitization

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Cited by 33 publications
(29 citation statements)
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“…It seems clear that this methodology could be applied with a high probability of success to conveniently conjugate meso-Cl near-IR dyes to antibodies, monobodies, and nanobodies to form selective agents for optical imaging in vivo. Traditional conjugation techniques tend to require modification of the dye to include maleimide or succinimide functionality [25,31,32], but the method developed here circumvents that process.…”
Section: Discussionmentioning
confidence: 99%
See 1 more Smart Citation
“…It seems clear that this methodology could be applied with a high probability of success to conveniently conjugate meso-Cl near-IR dyes to antibodies, monobodies, and nanobodies to form selective agents for optical imaging in vivo. Traditional conjugation techniques tend to require modification of the dye to include maleimide or succinimide functionality [25,31,32], but the method developed here circumvents that process.…”
Section: Discussionmentioning
confidence: 99%
“…Competition study of MHI-148 with amino acids in aqueous buffer Blocking experiments were performed to be absolutely sure that the vimentin Cys was the reactive group for coupling to MHI-148 in aqueous buffer at 37 °C. Thus, maleimide-blocked protein[24,25] was formed by incubating vimentin with 6-maleimidohexanoic acid (6-MA; 18 h in…”
mentioning
confidence: 99%
“…The eIF5 mutants were expressed in BL21(DE3) Codon Plus cells (Stratagene) as described (32). The purified proteins were then fluorescently labeled with fluorescein-maleimide (34).…”
Section: Methodsmentioning
confidence: 99%
“…Next we generated various single cysteine mutants of Cys-lite eIF5 by introducing cysteines at non-conserved, surface-exposed positions in the NTD, CTD, and linker region using sitedirected mutagenesis (Table 1). These mutants were then expressed in Escherichia coli and purified, and the cysteine residues were labeled with fluorescein-maleimide (34). We also labeled the three naturally occurring cysteines at positions 6, 289, and 294 in variants containing each one as the only surfaceexposed cysteine (Table 1).…”
Section: The Ctt Of Eif1a and N-terminal Domain Of Eif5 Move Closer Tmentioning
confidence: 99%
“…In this approach, a free thiol group (i.e., cysteine) on the protein is reacted with a fluorophore that is derivatized with a maleimide group, forming a thioether bond. It is a selective reaction around pH 7.0, though above pH 8.0, primary amines are also labeled (Nanda & Lorsch, 2014).…”
Section: Strategic Planningmentioning
confidence: 99%