CS231222B1 - Method of detection of protein separated in polyacrylamide gel - Google Patents
Method of detection of protein separated in polyacrylamide gel Download PDFInfo
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- CS231222B1 CS231222B1 CS824934A CS493482A CS231222B1 CS 231222 B1 CS231222 B1 CS 231222B1 CS 824934 A CS824934 A CS 824934A CS 493482 A CS493482 A CS 493482A CS 231222 B1 CS231222 B1 CS 231222B1
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Abstract
Vynález spatjé do odboru analytických, biochemických metod. Rieši spdsob detekcie bielkovín dělených v polyakrylamidovom géli, pri ktorom sa postupuje tak, že elektroforeagramy sa detekujú nasýteným roztokom 1:2 kovokomplexného arofarbiva chrómu, kobaltu alebo železa bis(1-feny1-3-alky1-5- -hydroxyl pyrozolyl) dibenzoovej kyseliny vzorca I kde X je chróm, kobalt alebo železg v roztoku kyseliny octovej alebo trichlorovej.The invention is more in the field of analytical, biochemical methods. It solves the way of detection proteins divided in polyacrylamide gel, in which the electrophoresis is performed are detected with a saturated solution 1: 2 metal-complex chromium, cobalt colorant or iron bis (1-phenyl-3-alkyl-5- -hydroxyl pyrozolyl) dibenzoic acid of Formula I wherein X is chromium, cobalt or iron in solution acetic or trichloric acid.
Description
Vynález sa týká zpOsobu detekcie bielkovín dělených v polyakrylamldovom géli.The invention relates to a method for detecting proteins separated in a polyacrylamide gel.
V súčasnej době sa na detekciu bielkovín používá 4-amino-5-hydroxy-3-(paranitrofenyla zo)-6-(fenylazo-2,7) naftalendisulfonová kyselina, dvojsodná soT ako procentuálny roztok v roztoku kyseliny octovej, triohloroctovej alebo v iných rozpúSťadlách, čo je popísané v práčach monografického charakteru autora Davisa D. A., Disc electrophoresie II,Currently, 4-amino-5-hydroxy-3- (paranitrophenyl) -6- (phenylazo-2,7) naphthalenedisulfonic acid, disodium salt, is used as a percentage solution in acetic, tri-chloroacetic acid or other solvents for protein detection. , which is described in monographs by Davis DA, Disc electrophoresie II,
N 4 Acad. Sci, 1964 a autora Maurera H. R., Disc elektrophoresié Theorie und Praxix der diskontinnierlicher Polyacrylamidgel-Elektrophorese, Walter de Gruyter and Co., Berlín, ,968. Tieto farbivá vykazujú krátkodobú trvanlivost v rozsahu nlekoTkých dní.N 4 Acad. Sci, 1964 and by Maurer H. R., Theory of Electrophoresis, Theory and Practice of Discontinuous Polyacrylamide Gel-Electrophores, Walter de Gruyter and Co., Berlin, 968. These dyes exhibit a short shelf life in the range of light days.
Uvedené nedostatky sú odstránené u zpdsobu detekcie bielkovín dělených v polyakrylamidovom géli podTa vynálezu, ktorého podstatou je, že na elektroforeogramy sa detekujú nasýtenýa roztokom 1:2 kovokomplexného azofarblva chrómu, kobaltu alebo železa bis (1-fenyl-3-alkyl-5-hydroxylpyrozolyl) dibenzoovej kyseliny vzorca I ch3-The above drawbacks are eliminated in the method for detecting proteins separated in a polyacrylamide gel according to the invention, which is based on the fact that electrophorograms are detected saturated with a 1: 2 solution of a metal-complex azo complex of chromium, cobalt or iron bis (1-phenyl-3-alkyl-5-hydroxyol) dibenzoic acid of formula I ch 3 -
COWHAT
N=N zN = N z
nn
-CH,CH,
COWHAT
(1) kde X je chróm, kobalt alebo železo v roztoku kyseliny octovej alebo trlchlóroctovej.(1) wherein X is chromium, cobalt or iron in acetic or trichloroacetic acid solution.
Výhodou zpdsobu detekcie bielkovín dělených v polyakrylamldovom géli podTa vynálezu je, že trvanlivost - stálost farbiva v závislosti na Sase za běžného osvetlenia a teploty přesahuje dobu jedného roka. Spektrum poskytovaných farieb je výhodné pre vyhodnotenie / v elektroforeogramov na rdznych typoch denzitometrov pro rdznej vlnovej dlžke. Určitý stupen rozdielnosti v selektivitě farbenie jednotlivých komponentov bielkovín umožňuje študovat kvalitativně aspekty delenia bielkovín.An advantage of the method of detecting proteins separated in the polyacrylamide gel according to the invention is that the shelf life of the dye in dependence on Saxon under normal illumination and temperature exceeds one year. The spectrum of colors provided is advantageous for evaluating / in electropherograms on different types of densitometers for different wavelengths. A certain degree of difference in selectivity staining of individual protein components makes it possible to study qualitative aspects of protein division.
V ddsledku substitúcie chrómu a metylových skupin inými radikájmi ako je kobalt alebo železo vzniká Široké farebné spektrum. Farbivá tohoto typu sa úspeSne aplikovali na detekciu bielkovín dělených elektroforeticky v polyakrylamldovom géli, nakoTko v oblasti pH pod izoelektrický bod molekule bielkoviny je kladné nabitá, čím sa vytvárajú podmienky pre viezanie elektrostatickými amóniovými zložkami farbiva.Due to the substitution of chromium and methyl groups by other radicals such as cobalt or iron, a wide color spectrum is produced. Dyes of this type have been successfully applied to the detection of proteins resolved electrophoretically in a polyacrylamide gel, since at a pH below the isoelectric point of the protein molecule it is positively charged, thereby creating conditions for the electrostatic ammonium dye components to be introduced.
Predmet vynálezu je ilustrovaný na příklade prevedenia.The invention is illustrated by way of example.
Příklad prevedenia'Execution example '
Rastlinné bielkoviny, izolované zo zrna obilnin, boli delené elektroforetickou metodou v polyakrylamldovom géli. Na priloženom obrázku je znázorněné farebné spektrum aplikovaných preparátor, farbených nesýtených-roztokom farbiva v 7 %-nej kyselino octovej. Farbenie sa uskutečnilo tak, že polyakrylamidové gély sa preniesli do kadičiek a farbivom a nechali stát po dobu 12 hodin. Potom sa prebytok detekčného činidla zlial a zvyšok činidla sa odstránil viacnésobnou dekontamináciou roztokom 7 %-nej kyseliny octovej a destilovanou vodou. Použilo se 1:2 kovokomplexné aroforbivo. 3Plant proteins isolated from cereal grain were separated by polyacrylamide gel electrophoresis. The attached figure shows the color spectrum of the applied preparators, stained with a non-saturated dye solution in 7% acetic acid. Staining was performed by transferring the polyacrylamide gels into beakers and dye and standing for 12 hours. Then the excess detection reagent was decanted and the remainder of the reagent was removed by repeated decontamination with a 7% acetic acid solution and distilled water. A 1: 2 metal-complex aroforbine was used. 3
Spčsob podTa vynálezu je možné využit okrem naznačeného směru na riešenie problemati ky stanovenia Specifických bielkovín, včítane enzýmov.The method according to the invention can be used in addition to the indicated direction to solve the problem of the determination of specific proteins, including enzymes.
Claims (1)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS824934A CS231222B1 (en) | 1982-06-30 | 1982-06-30 | Method of detection of protein separated in polyacrylamide gel |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS824934A CS231222B1 (en) | 1982-06-30 | 1982-06-30 | Method of detection of protein separated in polyacrylamide gel |
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| Publication Number | Publication Date |
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| CS493482A1 CS493482A1 (en) | 1984-02-13 |
| CS231222B1 true CS231222B1 (en) | 1984-10-15 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS824934A CS231222B1 (en) | 1982-06-30 | 1982-06-30 | Method of detection of protein separated in polyacrylamide gel |
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|---|---|
| CS (1) | CS231222B1 (en) |
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1982
- 1982-06-30 CS CS824934A patent/CS231222B1/en unknown
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| CS493482A1 (en) | 1984-02-13 |
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