JP2014077811A5 - - Google Patents

Download PDF

Info

Publication number
JP2014077811A5
JP2014077811A5 JP2014021163A JP2014021163A JP2014077811A5 JP 2014077811 A5 JP2014077811 A5 JP 2014077811A5 JP 2014021163 A JP2014021163 A JP 2014021163A JP 2014021163 A JP2014021163 A JP 2014021163A JP 2014077811 A5 JP2014077811 A5 JP 2014077811A5
Authority
JP
Japan
Prior art keywords
gel
dimensional
dimensional electrophoresis
satisfies
electrophoresis method
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
JP2014021163A
Other languages
Japanese (ja)
Other versions
JP2014077811A (en
JP5847213B2 (en
Filing date
Publication date
Application filed filed Critical
Priority to JP2014021163A priority Critical patent/JP5847213B2/en
Priority claimed from JP2014021163A external-priority patent/JP5847213B2/en
Publication of JP2014077811A publication Critical patent/JP2014077811A/en
Publication of JP2014077811A5 publication Critical patent/JP2014077811A5/ja
Application granted granted Critical
Publication of JP5847213B2 publication Critical patent/JP5847213B2/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Claims (4)

下記（１）の条件を満たす１次元目の等電点電気泳動を行う工程と、下記（２）の条件を満たす平衡化緩衝液処理を行う工程と、下記（３）の条件を満たす２次元目のＳＤＳ−ＰＡＧＥを行う工程と、を含み、
これらの各工程は、１次元目の等電点電気泳動、平衡化緩衝液処理、２次元目のＳＤＳ−ＰＡＧＥの順で行われることを特徴とする２次元電気泳動方法。
（１）１次元目電気泳動ゲルとして、その長手方向のゲル長が５〜１０ｃｍであって、２次元目電気泳動ゲルの検体適用側にそなえることができる扁平な棒状の形態を備えたものを用いる。
（２）上記等電点電気泳動を行ったゲルを、還元剤を含む緩衝液で平衡化し、及び、アルキル化剤を含む緩衝液で平衡化する。
（３）泳動方向基端部のゲル濃度を３〜６％とし、泳動方向先端側の部分のゲル濃度を１０〜２０％とした濃度勾配を有する２次元目電気泳動ゲルであるＢｉｓ−Ｔｒｉｓ Ｇｅｌを使用する。 The first dimension isoelectric focusing that satisfies the following condition (1), the equilibration buffer treatment that satisfies the following condition (2), and the two-dimensional condition that satisfies the following condition (3): Performing SDS-PAGE of the eye,
Each of these steps is performed in the order of first-dimensional isoelectric focusing, equilibration buffer treatment, and second-dimensional SDS-PAGE .
(1) as first dimensional electrophoresis gel, a longitudinal direction of the gel length of that is 5 to 10 cm, one with a form of flat bar-like that can be equipped on the sample application side of the second dimensional electrophoresis gel Is used.
(2) The gel subjected to the isoelectric focusing is equilibrated with a buffer containing a reducing agent and equilibrated with a buffer containing an alkylating agent.
(3) Bis-Tris Gel which is a second-dimensional electrophoresis gel having a concentration gradient in which the gel concentration at the base end in the migration direction is 3 to 6% and the gel concentration at the tip side in the migration direction is 10 to 20%. Is used. 検体が蛋白質を含むものであることを特徴とする、請求項１に記載の２次元電気泳動方法。2. The two-dimensional electrophoresis method according to claim 1, wherein the specimen contains a protein. 検体が、動物、植物、微生物由来の抽出物であり、かつ、蛋白質を含むものであることを特徴とする、請求項１に記載の２次元電気泳動方法。2. The two-dimensional electrophoresis method according to claim 1, wherein the specimen is an extract derived from an animal, a plant, or a microorganism and contains a protein. 更に、前記２次元電気泳動方法の検体の調製段階において、酸による沈殿及び有機溶媒による沈殿を行うことを特徴とする、請求項２又は請求項３に記載の２次元電気泳動方法。The two-dimensional electrophoresis method according to claim 2, further comprising performing precipitation with an acid and precipitation with an organic solvent in a specimen preparation stage of the two-dimensional electrophoresis method.
JP2014021163A 2014-02-06 2014-02-06 Two-dimensional electrophoresis method Active JP5847213B2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2014021163A JP5847213B2 (en) 2014-02-06 2014-02-06 Two-dimensional electrophoresis method

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP2014021163A JP5847213B2 (en) 2014-02-06 2014-02-06 Two-dimensional electrophoresis method

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
JP2009181812A Division JP5475358B2 (en) 2009-08-04 2009-08-04 Two-dimensional electrophoresis method

Publications (3)

Publication Number Publication Date
JP2014077811A JP2014077811A (en) 2014-05-01
JP2014077811A5 true JP2014077811A5 (en) 2014-06-19
JP5847213B2 JP5847213B2 (en) 2016-01-20

Family

ID=50783178

Family Applications (1)

Application Number Title Priority Date Filing Date
JP2014021163A Active JP5847213B2 (en) 2014-02-06 2014-02-06 Two-dimensional electrophoresis method

Country Status (1)

Country Link
JP (1) JP5847213B2 (en)

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS58140634A (en) * 1982-02-17 1983-08-20 Hitachi Ltd Method for simple two-dimentional electrophoresis
AUPO403496A0 (en) * 1996-12-05 1997-01-02 Macquarie Research Limited Electrophoresis separation methods
AUPR522501A0 (en) * 2001-05-25 2001-06-14 Proteome Systems Ltd Increased solubilisation of hydrophobic proteins
JP2005069935A (en) * 2003-08-26 2005-03-17 Hokkaido Green Bio Kenkyusho:Kk Variety determination method for raw wheat for wheat flour
DE112005003114T5 (en) * 2004-12-06 2008-05-08 Katayanagi Institute, Hachioji Method of separating protein, method of staining protein and liquid protein stain, and protein staining kit for use in these methods
JP2006294014A (en) * 2005-03-16 2006-10-26 Kumamoto Technology & Industry Foundation Analysis program, protein chip, method for manufacturing protein chip and antibody cocktail
US20070249014A1 (en) * 2006-02-10 2007-10-25 Invitrogen Corporation Labeling and detection of post translationally modified proteins
JP2008020383A (en) * 2006-07-14 2008-01-31 Sapporo Medical Univ Analysis method of humor protein using liposome as ligand and method for preparing humor protein
JP2009042004A (en) * 2007-08-07 2009-02-26 Norio Okuyama Electrophoretic support

Similar Documents

Publication Publication Date Title
EA201890945A1 (en) BIOLOGICAL PREPARATIONS AND THEIR APPLICATION WITH RESPECT TO PLANTS
JP2014000092A5 (en)
EP4219744A3 (en) Methods of preparing nucleic acids for sequencing
WO2015195949A3 (en) Methods, compositions, and devices for rapid analysis of biological markers
CN106415670A8 (en) For the supporting construction of the increasing material manufacturing of physical model
RU2020124757A (en) SYSTEM AND METHOD FOR COLLECTING NUCLEIC ACID SAMPLE
CY1123403T1 (en) MODIFIED HOST CELLS AND USES THEREOF
AR096236A1 (en) VHH SIMPLE DOMAIN ANTIBODIES DIRECTED TO NOROVIRUS GI.1 AND GII.4 AND ITS USES
GB2554572A (en) Physical linkage preservation in DNA storage
JP2018108063A5 (en)
JP2014209129A5 (en)
EA201590164A1 (en) PLANTS BRASSICA WITH MODIFIED SEEDS OIL COMPOSITION
JP2014077811A5 (en)
RU2014124569A (en) METHOD FOR NUCLEIC ACID ISOLATION
HOGAN Field Venom Extractions: Saving Fingers with Tubes, Forceps, and Nerf Bullets.
JP2013194009A5 (en)
MY170201A (en) A method for identifying a ganoderma-infected palm tree
RU2016141714A (en) A method of processing hatching eggs
EA201890136A1 (en) NUCLEIC ACID MOLECULE TO ENSURE INSECTICIDAL PROPERTIES IN PLANTS
Ghorbi et al. PHYSICOCHEMICAL METHODS IN ENDOTOXIN REMOVAL FROM RECOMBINANT ACTIVE PHARMACEUTICAL INGREDIENT (API)
RU2016124876A (en) The method for determining the elasticity of biological tissues
KR20170001332U (en) How to Sequence Proteins Binding to Hair
Cornejo Portugal et al. You ran without knowing where you were going" Migratory process of Yucatecan Mayans to San Francisco, California
TH1801000954A (en) Vaccines for treating HPV18
TH164822A (en) Methods and kits for optimum contour analysis