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Quantum dot bio-conjugate: as a western blot probe for highly sensitive detection of cellular proteins
Authors:Sonia Kale   Anup Kale   Haribhau Gholap   Abhimanyu Rana   Rama Desai   Arun Banpurkar   Satishchandra Ogale  Padma Shastry
Affiliation:(1) National Centre for Cell Science, Ganeshkhind, Pune, 411007, India;(2) Present address: Agharkar Research Institute, Pune, India;(3) Physical & Materials Chemistry Division, National Chemical Laboratory, Dr. Homi Bhabha Road, Pashan, Pune, 411008, India;(4) Present address: Center for Materials for Information Technology, University of Alabama, Tuscaloosa, 35487, AL, USA;(5) Department of Physics, University of Pune, Pune, 411007, India;
Abstract:In the present study, we report a quantum dot (QD)-tailored western blot analysis for a sensitive, rapid and flexible detection of the nuclear and cytoplasmic proteins. Highly luminescent CdTe and (CdTe)ZnS QDs are synthesized by aqueous method. High resolution transmission electron microscopy, Raman spectroscopy, fourier transform infrared spectroscopy, fluorescence spectroscopy and X-ray diffraction are used to characterize the properties of the quantum dots. The QDs are functionalized with antibodies of prostate apoptosis response-4 (Par-4), poly(ADP-ribose) polymerases and β actin to specifically bind with the proteins localized in the nucleus and cytoplasm of the cells, respectively. The QD-conjugated antibodies are used to overcome the limitations of conventional western blot technique. The sensitivity and rapidity of protein detection in QD-based approach is very high, with detection limits up to 10 pg of protein. In addition, these labels provide the capability of enhanced identification and localization of marker proteins in intact cells by confocal laser scanning microscopy.
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