Riken Corporation

Medical Biological Laboratories Co., Ltd.

Olympus Corporation

Japan Science and Technology Agency

The Institute of Physical and Chemical Research

Hamamatsu-photonics K.k.

Yokogawa Electric Corporation

Kobe University

Tokyo Metropolitan Institute of Medical Science

Atsushi Miyawaki

Satoshi Karasawa

Ryoko Ando

Takeharu Nagai

Takashi Fukano

Hideaki Mizuno

Hidekazu Tsutsui

Hiroshi Hama

Yasushi Aono

Toshio Araki

Masahiko Hirano

Masataka Kinjo

Kenta Saito

Hiroyuki Katayama

Takako Kogure

Asako Sawano

Akiko Kumagai

Takeo Tanaami

Yoshihiro Kawano

Kenta Mikuriya

Keiji Shimizu

Masafumi Oshiro

Naoki Seki

Naoshi Dohmae

Kiyochika Ishibashi

Kazuki Sasaki

Sachiko Karaki

Hisao Masai

Kiminori Ushida

Hiroyuki Imabayashi

Asako Sakaue

Hidetomo Tanase

Ichiro Morioka

Satoshi Iwano

Hajime Nakamura

Sota Iwatani

Akiko Masuda

Hidemitsu Furukawa

Hiroshi Kurokawa

Hiroyuki Kawano

Ryoko Brain Science And Life TechResFound Ando

Satoshi Inalab Karasawa

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.

PH-responsive proteolysis probe

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.

Modified luciferase

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. In detection step (iv), the fluorescence of UnaG is detected from the decomposition product of the sample and from the unreacted sample. Then, the amount of UB is derived from the difference between the detected values.

Measurement method for unbound bilirubin in blood sample

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) %.

Clarifying reagent for biological materials and use thereof

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.

Antibody composition, kit for preparing antibody composition, and immunostaining method

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).

Polypeptide exhibiting fluorescent properties, and utilization of the same

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.

Method for measuring autophagy

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.

Method for detecting bilirubin using a fluorescent protein

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.

Probe reagent for measurement of proteolytic activity

The present invention relates to fluorescent or luminescent probe reagents for measuring oxidative stress in a cell or an organism. Examples of the probe reagents include: a fluorescent or luminescent protein and a marker protein; a fluorescent or luminescent protein, a marker protein, and a regulatory factor; or a fluorescent or luminescent protein, a marker protein, a cleavage sequence, and a regulatory factor. In the probe reagents, the marker protein makes it possible to detect the oxidative stress caused by reactive oxygen species and comprises a regulatory factor-binding site and a ubiquitin-binding site; and the regulatory factor is a protein making it possible to regulate degradation of the marker protein in response to the reactive oxygen species. The present invention also relates to a method of measuring oxidative stress in a cell or an organism, or a method of screening a substance which suppresses or promotes the oxidative stress in a cell or an organism by using the probe reagent.

Probe reagent for measuring oxidative stress

An object of the present invention is to provide a red or orange fluorescent protein, which is characterized in that the difference (stokes shift) between an excitation peak value (wavelength of maximum absorption) and a fluorescence peak value (wavelength of maximum fluorescence) is greatened, so that the maximum fluorescence can be obtained by the maximum excitation. The present invention provides a novel fluorescent protein monomerized by introducing a mutation into a florescent protein derived fromsp., and a novel chromoprotein and fluorescent protein derived from. sp.

Fluorescent protein

An object of the present invention is to provide a novel fluorescent protein derived from. The present invention provides a fluorescent protein derived fromhaving the following properties:

An object of the present invention is to provide a method for introducing a physiologically active substance such as a gene into a cell, which introduces a physiologically active substance such as any given gene into any given cell in a view under a microscope, while significantly reducing invasiveness to the cell, and a device used for the above method. The present invention provides a method for introducing a physiologically active substance into a cell, which comprises: allowing a physiologically active substance to attach around a needle having a diameter of 500 nm or less, provided that it is able to be inserted into a cell; and inserting the above-described needle into the cell; and a microinjection device for carrying out the aforementioned method.

Microinjection method and device

It is an object of the present invention to provide a novel fluorescent protein and a novel chromoprotein. The present invention provides a novel fluorescent protein derived fromsp.,sp. and, and a novel chromoprotein derived from

Fluorescent protein and chromoprotein

An object of an embodiment of the present invention is to provide a method with which it is possible to easily distinguish a proliferation phase of a cell cycle from a resting phase thereof in real time. The object of the embodiment of the present invention is attained by providing a method for performing phase identification of the cell cycle, the method including: visualizing one or more gene-expression products as markers whose amounts in a cell change in a cell-cycle dependent manner; and detecting the products so as to distinguish the proliferation phase of the cell cycle from the resting phase thereof.

Probe for visualizing cell-cycle

Growing community of inventors

Saitama, Japan

Atsushi Miyawaki