| Budget Amount *help |
¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2018: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2017: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2016: ¥8,580,000 (Direct Cost: ¥6,600,000、Indirect Cost: ¥1,980,000)
|
|---|
| Outline of Final Research Achievements |
In this study, we aimed to develop a new enzymatic sensitization method for the sensitive detection of antigens on the cell surface. This method (CARP) is characterized by the use of hydrolytic enzymes, and the enzymatic reaction removes the hydrophilic part of the fluorescent substrate to become hydrophobic and stain the cytoplasm. The following results were obtained: 1) The CARP method was extended to β-galactosidase, which simplifies the detection procedures than the conventional alkaline phosphatase; 2) A covalent-binding substrate (the CLAMP method) was developed to inhibit the desorption of fluorescent molecules from cells, which is a problem of the CARP method. This was a molecule containing quinone methide. After the enzymatic reaction, the fluorescent substrate penetrated into the cell and covalently bound to the protein.
|
