Semiquantitative fluorescence method for bioconjugation analysis

Methods Mol Biol. 2014:1199:103-10. doi: 10.1007/978-1-4939-1280-3_8.

Abstract

Quantum dots (QDs) have been used as fluorescent probes in biological and medical fields such as bioimaging, bioanalytical, and immunofluorescence assays. For these applications, it is important to characterize the QD-protein bioconjugates. This chapter provides details on a versatile method to confirm quantum dot-protein conjugation including the required materials and instrumentation in order to perform the step-by-step semiquantitative analysis of the bioconjugation efficiency by using fluorescence plate readings. Although the protocols to confirm the QD-protein attachment shown here were developed for CdTe QDs coated with specific ligands and proteins, the principles are the same for other QDs-protein bioconjugates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cadmium Compounds / chemistry
  • Cattle
  • Humans
  • Immobilized Proteins / chemistry
  • Immunoglobulins / chemistry*
  • Quantum Dots / chemistry*
  • Serum Albumin, Bovine / chemistry*
  • Spectrometry, Fluorescence / methods*
  • Tellurium / chemistry
  • Water / chemistry

Substances

  • Cadmium Compounds
  • Immobilized Proteins
  • Immunoglobulins
  • Water
  • Serum Albumin, Bovine
  • Tellurium
  • cadmium telluride