Several automated methods have been reported to reduce the
amount of reagents needed for conducting WB.8−13 A
semiautomatic protein detection system was demonstrated
using a vacuum to actively drive the blocking reagent and
antibody solution through the membrane. This method can
complete WB in 30 min but consumes more energy without
reducing the required amount of primary antibody.9 Another
method used capillary isoelectric focusing to separate protein
samples, which were subsequently immobilized by photoactivated cross-linking to the capillary surface for detection by
flowing specific antibodies and reagents through the capillary.10
In addition to improved sensitivity and automation, this
method also provided better experimental reproducibility, but
the system is quite expensive to implement and does not
provide the size-based separation available in conventional WB.
Recently, a method named capillary electrophoresis-Western
blot (CEWB) has been shown to efficiently separate samples
based on their size-to-charge ratios in a small capillary. CEWB
eliminates the electro-blotting step, but the approach needs
further improvements in terms of robustness, throughput, and
detection limit. Moreover, CEWB implementation in biochemical laboratories is limited by its complicated apparatus
and procedures
Several automated methods have been reported to reduce theamount of reagents needed for conducting WB.8−13 Asemiautomatic protein detection system was demonstratedusing a vacuum to actively drive the blocking reagent andantibody solution through the membrane. This method cancomplete WB in 30 min but consumes more energy withoutreducing the required amount of primary antibody.9 Anothermethod used capillary isoelectric focusing to separate proteinsamples, which were subsequently immobilized by photoactivated cross-linking to the capillary surface for detection byflowing specific antibodies and reagents through the capillary.10In addition to improved sensitivity and automation, thismethod also provided better experimental reproducibility, butthe system is quite expensive to implement and does notprovide the size-based separation available in conventional WB.Recently, a method named capillary electrophoresis-Westernblot (CEWB) has been shown to efficiently separate samplesbased on their size-to-charge ratios in a small capillary. CEWBeliminates the electro-blotting step, but the approach needsfurther improvements in terms of robustness, throughput, anddetection limit. Moreover, CEWB implementation in biochemical laboratories is limited by its complicated apparatusand procedures
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