Abstract: A novel fluorescent protein and uses thereof are provided. A polypeptide of the present invention has a fluorescence property, the polypeptide being indicated in any one of (1) to (3) below: (1) a polypeptide having an amino acid sequence set forth in SEQ ID NO: 1; (2) a polypeptide having an amino acid sequence set forth in SEQ ID NO: 1, the amino acid sequence having replaced, deleted, inserted, and/or added therein not less than 1 and not more than 32 amino acids; and (3) a polypeptide having not less than 85% sequence identity with respect to an amino acid sequence set forth in SEQ ID NO: 1.

Abstract: A clearing reagent in accordance with an embodiment of the present invention for making a biological material transparent is a solution containing: at least one compound selected from the group consisting of urea and a urea derivative; sorbitol; and a surfactant which is contained at a concentration of 5 (w/v) % or less.

Abstract: A fluorescent protein of the present invention has an amino acid sequence of a green fluorescent protein (GFP) derived from a crystal jelly or of a mutant fluorescent protein of the green fluorescent protein, the amino acid sequence having an amino acid residue (alanine residue) substituted with a phenylalanine residue, the amino acid residue corresponding to position 206 with the amino acid sequence of the GFP used as a reference sequence.

Abstract: The present invention provides a tool which exhibits excellent properties in the quantification of autophagy activity. A unimolecular FRET probe of the present invention includes an acceptor consisting of a fluorescent protein to be enzymatically degraded inside a lysosome or a vacuole; and a donor having an amino acid sequence having a sequence identity of 95% or more with respect to an amino acid sequence represented by SEQ ID NO: 1.

Abstract: A method for making a biological material transparent according to the present invention includes: a first permeation step of causing a first solution to permeate into a biological material, the first solution containing at least one compound selected from the group consisting of urea and urea derivatives at a predetermined concentration; and then a second permeation step of causing a second solution to permeate into the biological material, the second solution containing at least one compound selected from the group consisting of urea and urea derivatives at a concentration higher than the concentration of the at least one compound contained in the first solution.

Abstract: A clearing reagent according to this invention for making a biological material transparent is a solution containing, as an active component, at least one compound selected from the group consisting of urea and urea derivatives, in order to provide a novel clearing reagent for making a biological material transparent.

Abstract: In order to provide modified luciferase whose substrate specificity to at least one luminescent substrate (e.g., AkaLumine) other than D-luciferin has been improved as compared with to D-luciferin, modified luciferase according to an aspect of the present invention has a mutation at an amino acid corresponding to a 347th amino acid in an amino acid sequence represented by SEQ ID NO: 1.

Abstract: Provided is a measurement method whereby the amount of unbound bilirubin (UB) can be exactly reflected whether a specimen contains a large amount of conjugated bilirubin or not. The measurement method for UB according to the present invention comprises decomposition step (i), decomposition stopping step (ii), contact step (iii) and detection step (iv). In decomposition step (i), a blood sample containing unconjugated bilirubin (iD-Bil) and conjugated bilirubin (D-Bil) is subjected to an oxidative decomposition reaction of UB in iD-Bil and D-Bil. In decomposition stopping step (ii), the oxidative decomposition reaction is stopped to give a decomposition product of the sample. In contact step (iii), the decomposition product of the sample is contacted with UnaG that is capable of specifically binding to iD-Bil. Separately, an unreacted sample, which is the blood sample not subjected to decomposition step (i), is contacted with UnaG too.

Abstract: A clearing reagent according to the present invention for making a biological material transparent is a solution containing: at a concentration of 1M or more and not more than 8.5M, at least one compound selected from the group consisting of urea and urea derivatives; and glycerol at a concentration of 25 (w/v) % or more and not more than 35 (w/v) %.

Abstract: In order to provide modified luciferase whose substrate specificity to at least one luminescent substrate (e.g., AkaLumine) other than D-luciferin has been improved as compared with to D-luciferin, modified luciferase according to an aspect of the present invention has a mutation at an amino acid corresponding to a 347th amino acid in an amino acid sequence represented by SEQ ID NO: 1.

Abstract: An antibody composition, which is an aspect of the present invention, contains at least one compound selected from the group consisting of urea and urea derivatives, the compound being contained in the antibody composition at a concentration of not less than 0.1 M and less than 1 M, the antibody composition being a solution.

Abstract: The present invention provides a tool which exhibits excellent properties in the quantification of autophagy activity. A unimolecular FRET probe of the present invention includes an acceptor consisting of a fluorescent protein to be enzymatically degraded inside a lysosome or a vacuole; and a donor having an amino acid sequence having a sequence identity of 95% or more with respect to an amino acid sequence represented by SEQ ID NO: 1.

Abstract: A mode of a polypeptide according to the present invention exhibits a fluorescence property, and has (1) an amino acid sequence represented by SEQ ID NO. 1 or NO. 2, (2) an amino acid sequence represented by SEQ ID NO. 1 or NO. 2 in which amino acid sequence 1 to 34 amino acids have been replaced or otherwise modified, (3) a sequence identity of 85% or more with respect to the amino acid sequence represented by SEQ ID NO. 1 or NO. 2, or (4) an amino acid sequence encoded by a polynucleotide that hybridizes under a stringent condition with a polynucleotide having a sequence complementary to a polynucleotide that encodes the polypeptide defined in (1).

Abstract: This invention relates to a method for measuring autophagy in cells, comprising using, as a probe reagent, a single fluorescent protein, to measure a change in fluorescence properties of the fluorescent probe reagent depending on pH changes associated with autophagy, thereby determining the presence or activity of autophagy, wherein the single fluorescent protein is resistant to degrading enzyme activity in the lysosome or vacuole of the cell, it is not denatured or inactivated under acidic to neutral pH environment, and it is capable of changing excitation spectra or fluorescence spectra when located under the environments of acidic region and neutral region.

Abstract: In order to provide a novel fluorescent protein and use thereof, the polypeptide according to the present invention has fluorescent properties in the presence of bilirubin and includes (1) the amino acid sequence of SEQ ID NO: 1, (2) an amino acid sequence having, for example, substitution of 1 to 21 amine acids in the amino acid sequence of SEQ ID NO: 1, (3) an amino acid sequence having 85% or more sequence identity to the amino acid sequence of SEQ ID NO: 1, or (4) the amino acid sequence encoded by a polynucleotide that hybridizes with a polynucleotide consisting of a sequence complementary to the polynucleotide encoding the polypeptide according to the amino acid sequence in (1) under a stringent condition.

Abstract: Provided is a measurement method whereby the amount of unbound bilirubin (UB) can be exactly reflected whether a specimen contains a large amount of conjugated bilirubin or not. The measurement method for UB according to the present invention comprises decomposition step (i), decomposition stopping step (ii), contact step (iii) and detection step (iv). In decomposition step (i), a blood sample containing unconjugated bilirubin (iD-Bil) and conjugated bilirubin (D-Bil) is subjected to an oxidative decomposition reaction of UB in iD-Bil and D-Bil. In decomposition stopping step (ii), the oxidative decomposition reaction is stopped to give a decomposition product of the sample. In contact step (iii), the decomposition product of the sample is contacted with UnaG that is capable of specifically binding to iD-Bil. Separately, an unreacted sample, which is the blood sample not subjected to decomposition step (i), is contacted with UnaG too.

Abstract: A clearing reagent in accordance with an embodiment of the present invention for making a biological material transparent is a solution containing: at least one compound selected from the group consisting of urea and a urea derivative; sorbitol; and a surfactant which is contained at a concentration of 5 (w/v) % or less.

Abstract: A mode of a polypeptide according to the present invention exhibits a fluorescence property, and has (1) an amino acid sequence represented by SEQ ID NO. 1 or NO. 2, (2) an amino acid sequence represented by SEQ ID NO. 1 or NO. 2 in which amino acid sequence 1 to 34 amino acids have been replaced or otherwise modified, (3) a sequence identity of 85% or more with respect to the amino acid sequence represented by SEQ ID NO. 1 or NO. 2, or (4) an amino acid sequence encoded by a polynucleotide that hybridizes under a stringent condition with a polynucleotide having a sequence complementary to a polynucleotide that encodes the polypeptide defined in (1).

Abstract: This invention relates to a probe reagent comprising, in order from the N-terminus to the C-terminus, the amino acid sequences of a fluorescent protein I, a peptide capable of terminating protein degradation (i.e., a degradation-terminating peptide), a spacer peptide, a fluorescent protein II, and a protein to be degraded, wherein the protein to be degraded is a protein degraded by the ubiquitin-proteasome system, and the probe reagent is degraded from the C-terminus, but that the degradation of the probe reagent is terminated at the degradation-terminating peptide, a nucleic acid encoding the probe reagent, and use of the probe reagent or the nucleic acid.

Abstract: In order to provide a novel fluorescent protein and use thereof, the polypeptide according to the present invention has fluorescent properties in the presence of bilirubin and includes (1) the amino acid sequence of SEQ ID NO: 1, (2) an amino acid sequence having, for example, substitution of 1 to 21 amine acids in the amino acid sequence of SEQ ID NO: 1, (3) an amino acid sequence having 85% or more sequence identity to the amino acid sequence of SEQ ID NO: 1, or (4) the amino acid sequence encoded by a polynucleotide that hybridizes with a polynucleotide consisting of a sequence complementary to the polynucleotide encoding the polypeptide according to the amino acid sequence in (1) under a stringent condition.