JPH0225745A - Protein molecular weight marker - Google Patents
Protein molecular weight markerInfo
- Publication number
- JPH0225745A JPH0225745A JP63175893A JP17589388A JPH0225745A JP H0225745 A JPH0225745 A JP H0225745A JP 63175893 A JP63175893 A JP 63175893A JP 17589388 A JP17589388 A JP 17589388A JP H0225745 A JPH0225745 A JP H0225745A
- Authority
- JP
- Japan
- Prior art keywords
- molecular weight
- protein
- weight marker
- marker
- protein molecular
- 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.)
- Pending
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
（技術分野）
この発明は、タンパク質分子量マーカーに関するもので
ある。さらに詳しくは、この発明は、生化学、医学、薬
学等の研究開発、あるいは医療診断等において有用な、
調製が容易で検定上ら有効な新しいタンパク質分子量マ
ーカーに関するものである。DETAILED DESCRIPTION OF THE INVENTION (Technical Field) This invention relates to protein molecular weight markers. More specifically, this invention is useful in research and development in biochemistry, medicine, pharmacy, etc., or in medical diagnosis, etc.
The present invention relates to a new protein molecular weight marker that is easy to prepare and effective in assay.
（従来の技術とその課題）
従来より、生化学、医学、薬学等の研究開発や、医療診
断においてタンパク質の分子量の概略値を検出すること
がしばしば行われており、このようなタンパク質分子量
の概略値の検出のための、分子量既知のタンパク質混合
物を標準試料として用いたＳＤＳ電気泳動法やゲルろ過
クロマトグラフィー法などが広く知られてもいる。(Prior art and its issues) Conventionally, it has often been necessary to detect approximate values of protein molecular weights in research and development in biochemistry, medicine, pharmacy, etc., and in medical diagnosis. SDS electrophoresis and gel filtration chromatography using a protein mixture of known molecular weight as a standard sample are widely known for detecting the value.
分子量の概略値を知るためのこの従来の方法は、種々の
限定条件はあるものの実用上有効なものとされてもいる
。This conventional method for determining the approximate value of molecular weight is said to be practically effective, although there are various limitations.
しかしながら、この従来の方法においては分子量の異な
るタンパク質試料を別個に採取し、これを混合すること
によって調製した標準試料を用いることが不可欠であり
、この際の調製には、（ア）個別のタンパク質試料を採
取しなければならないこと、（イ）個別のタンパク質試
料を各々別個に精製しなければならないこと、さらには
、（つ）これら各々のタンパク質試料の濃度をそろえて
混合しなければならないことから、非常に手間がかかり
、調製が簡単ではないという欠点がある。However, in this conventional method, it is essential to collect protein samples with different molecular weights separately and use a standard sample prepared by mixing them. (1) each individual protein sample must be purified separately; and (1) each protein sample must be mixed to the same concentration. However, it has the disadvantage that it is very time-consuming and not easy to prepare.
この発明は、以上の通りの事情に鑑みてなされたもので
あり、従来の標準試料の調製法の欠点を解消し、調製が
容易で分子量の検定に有効に用いることのできる新しい
タンパク質分子量マーカーその調製法を提供することを
目的としている。This invention was made in view of the above circumstances, and it provides a new protein molecular weight marker that overcomes the drawbacks of conventional standard sample preparation methods, is easy to prepare, and can be effectively used for molecular weight assay. The purpose is to provide a method of preparation.
（課題を解決するための手段）
この発明は、上記の課題を解決するために、タンパク質
試料を化学的または酵素的に切断して得られる断片化反
応生成物からなることを特徴とするタンパク質分子量マ
ーカーを提供するものである。(Means for Solving the Problems) In order to solve the above-mentioned problems, the present invention provides a protein molecular weight fragmentation product comprising a fragmentation reaction product obtained by chemically or enzymatically cleaving a protein sample. It provides a marker.
また、この発明は、このタンパク質分子量マーカーの調
製法をも提供する。The invention also provides a method for preparing this protein molecular weight marker.
切断反応は、対象とするタンパク質試料を適宜な化学的
または酵素切断剤を用いることによって行うことができ
る。たとえば、次の表−１および表−２に示したような
化学的切断剤もしくはプロテアーゼを用いることができ
る。この切断反応によって得られる断片化した反応生成
物をタンパク質分子量マーカーとする。The cleavage reaction can be carried out by treating the target protein sample with an appropriate chemical or enzymatic cleavage agent. For example, chemical cleavage agents or proteases such as those shown in Tables 1 and 2 below can be used. The fragmented reaction product obtained by this cleavage reaction is used as a protein molecular weight marker.
表−１
表−２
（作 用）
化学的または酵素的に切断した反応生成物を分子量マー
カーとするため、従来の標準試料のように、分子量の相
異する各々の試料を個別に採取、精製する必要はなく、
また、濃度の調節も必要ではない、このため、分子量マ
ーカーとしての調製は極めて容易であり、簡便に分子量
マーカーとして使用することができる。Table 1 Table 2 (Function) In order to use chemically or enzymatically cleaved reaction products as molecular weight markers, samples with different molecular weights are individually collected and purified, just like conventional standard samples. There is no need to
In addition, there is no need to adjust the concentration, so it is extremely easy to prepare as a molecular weight marker and can be easily used as a molecular weight marker.
（実施例）
以下、この発明の実施例を示す、もちろんこの発明は以
下の実施例によって限定されるものではない。(Examples) Examples of the present invention will be shown below, but of course the present invention is not limited to the following examples.
実施例としては、サルモネラ菌のべん毛繊維タンパク質
を当初の試料タンパク質とした。As an example, the flagellar fiber protein of Salmonella enterica was used as the initial sample protein.
このサルモネラ菌べん毛繊維タンパク質的１■に７０％
ギ酸２ｍｌを加えて溶解し、１０ｍ１のすり合わせ栓付
試験管に移しな、この中に、Ｂ ｒＣＮ（ブロモシアン
）の７０％ギ酸の溶液（１度６０ｑ／ａ＋ｌ ） ２０
０ μｍを加え、３７℃の温度で２日間反応させて断片
化反応を行った。この断片化反応によって得られた反応
混合物から減圧下に溶媒を留去して、分子量５１，００
０のサルモネラ閑べん毛繊維タンパク質から、次の分子
量、
４８．０００
３１ 、０００
１９．０００
１６．０００
の各々のタンパク質が得られた。70% of this salmonella flagellar fiber protein
Add 2 ml of formic acid to dissolve and transfer to a 10 ml test tube with a ground stopper.Into this, add a solution of 70% formic acid (60 q/a+l at a time) of B rCN (bromo cyanide) 20
A fragmentation reaction was performed by adding 0 μm and reacting at a temperature of 37° C. for 2 days. The solvent was distilled off under reduced pressure from the reaction mixture obtained by this fragmentation reaction, and the molecular weight was 51,000.
From 0 Salmonella flagellar fiber proteins, proteins with the following molecular weights: 48.000 31 , 000 19.000 16.000 were obtained.
これらのタンパク質について５ＤＳ−ポリアクリルアミ
ドゲル電気泳動により相対移動度と分子量との相関性を
測定した。これを示したものが第１図である。電気泳動
による移動距離と分子量との間には極めて良好な相関関
係が認められる。これを検定曲線とすることによって、
タンパク質の分子量を手軽に推定することができる。The correlation between relative mobility and molecular weight of these proteins was measured by 5DS-polyacrylamide gel electrophoresis. FIG. 1 shows this. An extremely good correlation is observed between electrophoretic migration distance and molecular weight. By using this as the test curve,
The molecular weight of proteins can be easily estimated.
（発明の効果）
この発明により、個別のタンパク質の採取、精製、さら
にこれらタンパク質の混合にとらなう濃度の調節も必要
でない、その調製の容易なタンパク質分子量マーカーが
得られる。(Effects of the Invention) According to the present invention, it is possible to obtain a protein molecular weight marker that is easy to prepare and does not require collection and purification of individual proteins, nor adjustment of concentration required for mixing these proteins.
タンパク質の分子量検定に有効なマーカーが実現される
。A marker effective for protein molecular weight assay is realized.
第１図は、この発明の一例について分子量と電気泳動の
相対位動距離との関係を示した相関図である。FIG. 1 is a correlation diagram showing the relationship between molecular weight and relative electrophoretic position distance for an example of the present invention.
Claims (2)
得られる断片化反応生成物からなることを特徴とするタ
ンパク質分子量マーカー。(1) A protein molecular weight marker comprising a fragmentation reaction product obtained by chemically or enzymatically cleaving a protein sample.
断片化反応生成物とすることを特徴とするタンパク質分
子量マーカーの調製法。(2) A method for preparing a protein molecular weight marker, which comprises cleaving a protein sample chemically or enzymatically to obtain a fragmentation reaction product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63175893A JPH0225745A (en) | 1988-07-14 | 1988-07-14 | Protein molecular weight marker |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63175893A JPH0225745A (en) | 1988-07-14 | 1988-07-14 | Protein molecular weight marker |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0225745A true JPH0225745A (en) | 1990-01-29 |
Family
ID=16004069
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63175893A Pending JPH0225745A (en) | 1988-07-14 | 1988-07-14 | Protein molecular weight marker |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0225745A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7776228B2 (en) | 2006-04-11 | 2010-08-17 | Ebara Corporation | Catalyst-aided chemical processing method |
| JP2011050272A (en) * | 2009-08-31 | 2011-03-17 | Univ Of Ryukyus | Molecular weight marker and method for preparing the same |
| EP2381008A2 (en) | 2006-08-28 | 2011-10-26 | Osaka University | Catalyst-aided chemical processing method and apparatus |
-
1988
- 1988-07-14 JP JP63175893A patent/JPH0225745A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7776228B2 (en) | 2006-04-11 | 2010-08-17 | Ebara Corporation | Catalyst-aided chemical processing method |
| US8679286B2 (en) | 2006-04-11 | 2014-03-25 | Ebara Corporation | Catalyst-aided chemical processing method |
| EP2381008A2 (en) | 2006-08-28 | 2011-10-26 | Osaka University | Catalyst-aided chemical processing method and apparatus |
| JP2011050272A (en) * | 2009-08-31 | 2011-03-17 | Univ Of Ryukyus | Molecular weight marker and method for preparing the same |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jordan | Studies on 5 s RNA conformation by partial ribonuclease hydrolysis | |
| Sanger | The terminal peptides of insulin | |
| US4287184A (en) | Process for healing wounds | |
| Müller et al. | Co-expression of the subunits of the heterodimer of HIV-1 reverse transcriptase in Escherichia coli | |
| Crawford et al. | Reaction of malonaldehyde with protein | |
| Kessler et al. | Type I procollagen C‐proteinase from mouse fibroblasts: Purification and demonstration of a 55‐kDa enhancer glycoprotein | |
| Machold et al. | Resolution of the light-harvesting chlorophyll ab-protein of Vicia faba chloroplasts into two different chlorophyll-protein complexes | |
| Budzko et al. | Cleavage of the fourth component of human complement (C4) by C1 esterase: isolation and characterization of the low molecular weight product | |
| Heimbach et al. | Affinity purification of the HIV-1 protease | |
| Myers | Studies on cholinesterase. 7. Determination of the molar concentration of pseudo-cholinesterase in serum | |
| Dostál et al. | Isolation and some chemical properties of aspermatogenic substance from bull seminal vesicle fluid | |
| Taniuchi et al. | Ligand-induced resistance of staphylococcal nuclease and nuclease-T to proteolysis by subtilisin, α-chymotrypsin, and thermolysin | |
| Beaven et al. | Nitration of ribonuclease | |
| Sokolovsky et al. | Reaction of tetranitromethane with tryptophan and related compounds | |
| Cory et al. | Isolation and characterization of enolase from rainbow trout (Salmo gairdnerii gairdnerii) | |
| Burrell et al. | Binding of ribosomal proteins to RNA covalently coupled to agarose | |
| Jameson et al. | Affinity chromatography of bovine trypsin. A rapid separation of bovine α-and β-trypsin | |
| Berns et al. | The molecular weight of the 14S calf lens messenger RNA | |
| Abdelrahim et al. | Liquid chromatographic assay of dityrosine in human cerebrospinal fluid | |
| Gordon et al. | α1 acute-phase globulins of rats. Isolation and some properties | |
| Louis et al. | Protein components of sarcoplasmic reticulum membranes from different animal species | |
| Nomoto et al. | Presence of nickel in alpha-2 macroglobulin isolated from human serum by high performance liquid chromatography | |
| Iannucci et al. | Purification of lumbrokinase from Eisenia fetida using aqueous two-phase systems and anion-exchange chromatography | |
| JPH01160496A (en) | Method for selectively degrading fusion proteins | |
| Wolosowicz et al. | Method for Determining ε-Aminocaproic Acid (EACA) in Blood Plasma, Based on the Antifibrinolytic Properties of this Compound |