Detection of proteins cross-linked within galactoside polyacrylate-based hydrogels by means of a quantum dot fluororeagent

Goldman, Ellen R.; O'Shaughnessy, Thomas J.; Soto, Carissa M.; Patterson Jr., Charles H.; Taitt, Chris R.; Spector, Mark S.; Charles, Paul T.
December 2004
Analytical & Bioanalytical Chemistry;Dec2004, Vol. 380 Issue 7/8, p880
Academic Journal
Protein toxins have been immobilized in a galactoside polyacrylate hydrogel in a microarray format. The large pore size and solution-like environment of these novel hydrogels allow for easy penetration of large proteins and detection reagents. Confocal microscopy provided three-dimensional visualization of dye-labeled toxins cross-linked within the gel and of streptavidin-coated quantum dot (QD) fluorophores used to visualize the toxins after incubation with biotinylated anti-toxin antibodies. Fluorescence microscopy was utilized to visualize arrays of toxins detected by a biotinylated antibody and then exposure to streptavidin-conjugated QDs. The intensity of the QD fluorescence was quantified, and binding to two toxins on three types of hydrogels was examined.


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