Hydrophobic adsorption chromatography to reduce nonspecific staining by rhodamine-labeled antibodies

Edward G. Spack, Beverly Packard, Marjorie L. Wier, Michael Edidin

Research output: Contribution to journalArticle

Abstract

Rhodamine adsorbed to protein was removed from rhodamine-conjugated antibody solutions by adsorption to hydrophobic macroporous beads (Bio-Beads SM-2) following gel permeation chromatography. This simple technique eliminated the contaminating free dye more effectively than gel filtration alone, but neither significantly reduced the level of fluorescently conjugated antibody nor altered the latter's binding characteristics. Passage of the fluorescent antibody solution over the SM-2 beads eliminated the high background of nonspecific staining caused by internalization of residual free dye.

Original languageEnglish (US)
Pages (from-to)233-237
Number of pages5
JournalAnalytical biochemistry
Volume158
Issue number1
DOIs
StatePublished - Oct 1986

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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