A novel antibody microarray format using non-covalent antibody immobilization with chemiluminescent detection.

To date, protein and antibody microarrays have been used in reverse-phase and sandwich-based methods in order to detect known proteins such as biomarkers in samples. Our group developed "libraries" of antibodies against unknown proteins, referred to as mKIAA proteins, and we attempted to discover candidate novel biomarkers by protein expression profiling.To profile mKIAA protein expression using these antibodies, we established an antibody microarray system using chemiluminescent detection.

A chip-based miniaturized format for protein-expression profiling: the exploitation of comprehensively produced antibodies.

Numerous antibodies have been developed and validated in recent years, and show promise for use in novel functional protein assays. Such assays would be an alternative to pre-existing comprehensive assays, such as DNA microarrays. Antibody microarrays are thought to represent those functional protein assays.

Targeted expression of Ip3 sponge and Ip3 dsRNA impaires sugar taste sensation in Drosophila.

We evaluated the role of IP(3) in sugar taste reception in Drosophila melanogaster by inactivating the IP(3) signaling using genetic tools. We used the "IP(3) sponge," composed of the modified ligand-binding domain from the mouse IP(3) receptor, which was designed to absorb IP(3) in competition with native IP(3) receptors. Another tool was a transgene that generates double-stranded RNA against IP(3) receptor mRNA.

Label-free detection of proteins in crude cell lysate with antibody arrays by a surface plasmon resonance imaging technique.

We established a label-free method of measuring proteins in crude cell lysate using antibody arrays and surface plasmon resonance (SPR) imaging. The refractivity of the running buffer was adjusted with that of the lysate to overcome the bulk effect. The chemistries of the fabricated arrays were investigated to reduce nonspecific adsorption on the array surface.

Species-specific patterns of sexual dimorphism in the expression of fruitless protein, a neural musculinizing factor in Drosophila.

In Drosophila melanogaster, male-specific forms of the fruitless (fru) gene product, mFru protein, function as a neural sex-determination factors that directs the development of at least two male characteristics, namely courtship and mating behavior and the formation of the muscle of Lawrence (MOL). In D. melanogaster, the male-specific expression of Fru protein in motoneurons is responsible for the male-limited induction of the MOL by such neurons.

A novel approach to protein expression profiling using antibody microarrays combined with surface plasmon resonance technology.

We have previously described our systems for the high-throughput production of antibodies against mouse KIAA proteins and their validation (Proteomics 2004, 4, 1412-1416). Using our "libraries" of antibodies, we established a novel antibody microarray system in which surface plasmon resonance (SPR) technology is utilized for signal detection. Up to 400 real-time antibody-target bindings could be measured simultaneously within a single hour.

Male-specific expression of the fruitless protein is not common to all Drosophila species.

Sex-specific behavioral patterns must be a result of sexual differences in the structure and/or function of the central nervous system (CNS). Male Drosophila melanogaster mutants for the fruitless (fru) locus exhibit enhanced male-to-male courtship. The fru mutant males are accompanied by malformation of the male-specific muscle of Lawrence (MOL), which, in wild-type males, is induced by male motoneurons innervating it.

Expression analysis of the lingerer gene in the larval central nervous system of Drosophila melanogaster.

The lingerer (lig) gene is necessary for initiation and termination of copulatory behavior in Drosophila melanogaster. The lig gene encodes cytoplasmic proteins, and is expressed in the central nervous system (CNS) during the late third-instar larval stage when the lig function is required for normal copulation to occur after adult eclosion. To characterize the lig-expressing cells in the late third-instar larval CNS, we have isolated a genomic fragment containing the promoter/enhancer region of the lig gene, and established transgenic lines in which expression of reporter genes is controlled by the lig promoter/enhancer.

Pathology of the adult central nervous system induced by genetic inhibition of programmed cell death in Drosophila pupae.

In the spinster (spin) mutant of Drosophila melanogaster, the extent of programmed cell death (PCD) in the abdominal ganglion 6 h after puparium formation (APF) is significantly reduced. The shortening of the abdominal ganglion, which is normally completed 48 h APF, does not occur. After eclosion, neurodegeneration accompanied by accumulation of autofluorescent materials is manifested in the central nervous system (CNS) of the spin mutant.