Laser-induced fluorescence detector for capillary-based isoelectric immunoblot assay.
We describe a whole-capillary, multicolor laser-induced fluorescence scanner for microfluidic protein analysis systems. Separation of proteins is achieved by isoelectric focusing in a short length of fused-silica capillary after which the resolved proteins are immobilized to the capillary wall using photochemistry. The capillary is then evacuated, and fluorescently labeled antibodies are flowed through the capillary to bind to the immobilized proteins. This technique provides high sensitivity, the ability to spatially resolve and quantify proteins, and provides the opportunity for complete automation. Results obtained by fluorescence detection are compared to those obtained by chemiluminescence while offering enhanced resolution and signal stability.
Knittle
James E
JE
Cell Biosciences, Inc., 1050 Page Mill Road, Palo Alto, California 94304, USA. jknittle@cellbiosciences.com
Roach
David
D
Horn
Peter B Vander
PB
Voss
Karl O
KO
D004867:Equipment Design; D005453:Fluorescence; D015151:Immunoblotting / Q000295:instrumentation / Q000379:methods*; D007525:Isoelectric Focusing; D007834:Lasers