TY - JOUR A1 - Grabolle, Markus A1 - Brehm, Robert A1 - Pauli, Jutta A1 - Dees, F.M. A1 - Hilger, I. A1 - Resch-Genger, Ute T1 - Determination of the labeling density of fluorophore-biomolecule conjugates with absorption spectroscopy JF - Bioconjugate chemistry N2 - Dye–biomolecule conjugation is frequently accompanied by considerable spectral changes of the dye’s absorption spectrum that limit the use of the common photometrical method for the determination of labeling densities. Here, we describe an improvement of this method using the integral absorbance of the dye instead of its absorbance at the long wavelength maximum to determine the concentration of the biomolecule-coupled dye. This approach is illustrated for three different cyanine dyes conjugated to the antibody IgG. KW - Fluorescent dye KW - Aggregation KW - Dimerization KW - Fluorophore-labeled antibodies KW - Labeling density KW - Dye-to-protein ratio KW - Absorption PY - 2012 DO - https://doi.org/10.1021/bc2003428 SN - 1043-1802 SN - 1520-4812 VL - 23 IS - 2 SP - 287 EP - 292 CY - Washington, DC AN - OPUS4-25527 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -