polyacrylamide Dotaz Zobrazit nápovědu
Heparin was coupled via its carboxyl group with a polyacrylamide derivative containing covalently bound amino groups using the carbodiimide reaction. Heparin immobilized in this way proved to be useful as an affinity carrier for the isolation of antithrombin III and heparin-binding proteins from boar seminal plasma.
- MeSH
- akrylové pryskyřice chemie MeSH
- antithrombin III izolace a purifikace MeSH
- chromatografie afinitní MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- heparin chemie MeSH
- karbodiimidy chemie MeSH
- prasata MeSH
- sperma chemie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- akrylové pryskyřice MeSH
- antithrombin III MeSH
- heparin MeSH
- karbodiimidy MeSH
- polyacrylamide gels MeSH Prohlížeč
The present review concentrates on techniques for the staining and quantification of proteins separated by polyacrylamide gel electrophoresis. Staining with organic dyes has been used for approximately thirty years; the silver staining technique was introduced in 1979. The problems of silver staining are presented separately because the mechanism of this staining is in principle different from staining with organic dyes. Less attention has been devoted to quantification of two-dimensional gels, because this autoradiography is preferred because of its high sensitivity and fewer problems with accurate quantification in contrast to silver staining.
- MeSH
- barvení a značení * metody MeSH
- elektroforéza v polyakrylamidovém gelu * MeSH
- proteiny izolace a purifikace MeSH
- stříbro MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- proteiny MeSH
- stříbro MeSH
The separation of oligonucleotides by capillary zone electrophoresis (CZE) was studied in fused silica separation capillaries filled by linear (noncrosslinked) polyacrylamide (PAA) solutions, introduced into the capillary from the stock by pressure after each analysis. The time-consuming in-capillary polymerization step could thus be avoided, and fast and reproducible repetition of the analyses was assured. The PAA concentrations varied within the range of 3-10% and both the reproducibility of the analyses and the stability of the solution in the capillary, with and without a chemically treated inner wall, were tested. Ferguson plots were used to assess the size selectivity of the separation.
- MeSH
- elektroforéza v polyakrylamidovém gelu metody MeSH
- oligonukleotidy izolace a purifikace MeSH
- reprodukovatelnost výsledků MeSH
- tlak MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- oligonukleotidy MeSH
Native polyacrylamide electrophoresis in the presence of two reversible protein anionic stains (Ponceau S and Ponceau 2R) was used to study the oligomeric states of soluble proteins. A mild binding of the used protein stains to nondissociated protein oligomers imposed a charge shift on the proteins resulting into separation of protein species according to their size under physiological conditions. Adsorbed stains could be easily removed after electrophoresis by washing of polyacrylamide gel with buffer and protein complexes could be visualized either by the detection of their enzyme activity or by using a nonspecific protein stain. The specific detection of enzyme activity of glycosidases, lactate dehydrogenase, or phosphatases was shown as an example.
- MeSH
- azosloučeniny chemie MeSH
- barvení a značení MeSH
- elektroforéza v polyakrylamidovém gelu metody MeSH
- multimerizace proteinu MeSH
- proteiny chemie MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Názvy látek
- azosloučeniny MeSH
- ponceau S MeSH Prohlížeč
- proteiny MeSH
In this work, we present a home-made two-dimensional (2-D) CCD imaging system for the monochromatic densitometry of plane gels and its application to the imaging and densitometry of chlorophyll (Chl)-containing proteins separated by non-denaturing polyacrylamide gel electrophoresis. The monochromatic imaging of separated green bands at the wavelengths corresponding to their absorption band increases their contrast. This allows a better visualization of the faint-green bands in the gel and using of samples with lower Chl content for the electrophoresis. By the comparison of 2-D densitograms of the same gel illuminated with 670 and 650 nm lights, that is, at the red absorption maximum of Chl a and b, respectively, we achieved a selective imaging of the complexes with different Chl a/b ratio. This approach was used to specify an unknown band that appeared in the gel of the sample prepared from the thylakoid membranes of preheated barley leaves.
- MeSH
- 2D gelová elektroforéza metody MeSH
- chlorofyl analýza MeSH
- elektroforéza v polyakrylamidovém gelu metody MeSH
- fotosyntetické reakční centrum - proteinové komplexy izolace a purifikace MeSH
- ječmen (rod) růst a vývoj MeSH
- světlosběrné proteinové komplexy MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- chlorofyl MeSH
- fotosyntetické reakční centrum - proteinové komplexy MeSH
- světlosběrné proteinové komplexy MeSH
Alkaline hydrolysis of high-molecular-mass (10 MDa) linear polyacrylamide, a widely used replaceable sieving medium for the capillary electrophoresis of DNA fragments and proteins, was investigated. The release rate of ammonia, a product of amide group hydrolysis, was monitored by a high-sensitivity continuous-flow system. The experimental results show a rapid onset of hydrolysis at a temperature of 70 degrees C. While the degree of 1 h hydrolysis was evaluated to reach about 16% at 70 degrees C and pH 13, it drops to 5% at 50 degrees C and pH 12.5.
- MeSH
- akrylové pryskyřice chemie MeSH
- DNA chemie MeSH
- elektroforéza kapilární přístrojové vybavení MeSH
- lidé MeSH
- proteiny chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- akrylové pryskyřice MeSH
- DNA MeSH
- polyacrylamide MeSH Prohlížeč
- proteiny MeSH
- MeSH
- akrylové pryskyřice chemie MeSH
- DNA fingerprinting metody MeSH
- katalýza MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- akrylové pryskyřice MeSH
- polyacrylamide gels MeSH Prohlížeč
Described is an alternative procedure for the phenotyping of pig alpha 1B-glycoprotein (PO2) and haemopexin. The procedure is based on the separation of serum samples by horizontal polyacrylamide gel electrophoresis, passive blotting onto a nitrocellulose (NC) sheet, and immunochemical detection using a mixture of a primary antibody (rabbit anti-pig alpha 1B or anti-pig haemopexin) and a peroxidase-labelled secondary antibody. Several NC copies can be obtained from a single gel and these can be developed with different monospecific antisera.
- MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- fenotyp MeSH
- hemopexin genetika MeSH
- imunoblotting MeSH
- imunoenzymatické techniky * MeSH
- krevní proteiny genetika MeSH
- prasata MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- hemopexin MeSH
- krevní proteiny MeSH
A modified method for the detection of DNA polymerases in cell extracts and purified enzyme preparations after electrophoresis in polyacrylamide gradient cylindrical gels is described. The technique, which is based on direct assay of activity in a reaction mixture during elution of DNA polymerases from gel slices, was applied to the pursuit of enzyme forms of Streptomyces aureofaciens DNA polymerase during purification procedure. In a crude extract of S. aureofaciens mycelium many catalytically active forms of DNA polymerase ranging from 35 to 860 kDa were detected. After purification, that included mycelium homogenization, precipitation of nucleic acids by polyethylene glycol, DEAE-Sephadex, QAE-Sephadex and DNA-Sepharose chromatography, higher molecular mass forms of more than 172 kDa have not been found. The lower molecular mass forms were separated into two groups by DNA-Sepharose chromatography. On the basis of their characterization, it is assumed that the lower molecular mass forms are produced by proteolysis and the major form found after purification of S. aureofaciens DNA polymerase in the presence of suitable proteinase inhibitors should be a protein of 172 kDa.
