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Magnetic bead assisted labeling of antibodies at nanogram scale
Author(s) -
Dezfouli Mahya,
Vickovic Sanja,
Iglesias Maria Jesus,
Nilsson Peter,
Schwenk Jochen M.,
Ahmadian Afshin
Publication year - 2014
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201300283
Subject(s) - chemistry , antibody , proteome , fluorescent labelling , chromatography , reagent , horseradish peroxidase , covalent bond , fluorescence , biochemistry , enzyme , biology , physics , organic chemistry , quantum mechanics , immunology
There are currently several initiatives that aim to produce binding reagents for proteome‐wide analysis. To enable protein detection, visualization, and target quantification, covalent coupling of reporter molecules to antibodies is essential. However, current labeling protocols recommend considerable amount of antibodies, require antibody purity and are not designed for automation. Given that small amounts of antibodies are often sufficient for downstream analysis, we developed a labeling protocol that combines purification and modification of antibodies at submicrogram quantities. With the support of magnetic microspheres, automated labeling of antibodies in parallel using biotin or fluorescent dyes was achieved.