The Experts below are selected from a list of 190356 Experts worldwide ranked by ideXlab platform
Bryan John Smith - One of the best experts on this subject based on the ideXlab platform.
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SDS Polyacrylamide Gel Electrophoresis of Proteins
Basic Protein and Peptide Protocols, 2003Co-Authors: John M. Walker, Bryan John SmithAbstract:INTRODUCTIONThis protocol describes the separation of proteins by SDS-Polyacrylamide Gel Electrophoresis. SDS is used with a reducing agent and heat to dissociate the proteins. SDS-polypeptide complexes form and migrate through the Gels according to the size of the polypeptide. By using markers of known molecular weight, the molecular weight of the polypeptide chain(s) can be estimated.
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SDS Polyacrylamide Gel Electrophoresis of Proteins.
Methods in molecular biology (Clifton N.J.), 1994Co-Authors: Bryan John SmithAbstract:Probably the most widely used of techniques for analyzing mixtures of proteins is SDS Polyacrylamide Gel Electrophoresis. In this technique, proteins are reacted with the anionic detergent, sodium dodecylsulfate (SDS, or sodium lauryl sulfate) to form negatively charged complexes. The amount of SDS bound by a protein, and so the charge on the complex, is roughly proportional to its size. Commonly, about 1.4 g SDS is bound per 1 g protein, although there are exceptions to this rule. The proteins are generally denatured and solubilized by their binding of SDS, and the complex forms a prolate elipsoid or rod of a length roughly proportionate to the protein's molecular weight. Thus, proteins of either acidic or basic pI form negatively charged complexes that can be separated on the bases of differences in charges and sizes by Electrophoresis through a sieve-like matr ix of Polyacrylamide Gel.
Joseph D. Puglisi - One of the best experts on this subject based on the ideXlab platform.
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RNA purification by preparative Polyacrylamide Gel Electrophoresis.
Methods in enzymology, 2013Co-Authors: Alexey Petrov, Elisabetta Viani Puglisi, Joseph D. PuglisiAbstract:Preparative Polyacrylamide Gel Electrophoresis (PAGE) is a powerful tool for purifying RNA samples. Denaturing PAGE allows separation of nucleic acids that differ by a single nucleotide in length. It is commonly used to separate and purify RNA species after in vitro transcription, to purify naturally occurring RNA variants such as tRNAs, to remove degradation products, and to purify labeled RNA species. To preserve RNA integrity following purification, RNA is usually visualized by UV shadowing or stained with ethidium bromide or SYBR green dyes.
Halina Porowska - One of the best experts on this subject based on the ideXlab platform.
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the use of preparative Polyacrylamide Gel Electrophoresis and electroelution for purification of mucus glycoproteins
Analytical Biochemistry, 1995Co-Authors: A Paszkiewiczgadek, Andrzej Gindzienski, Halina PorowskaAbstract:This paper describes a novel technique for purifying glycoproteins from porcine gastric mucus by preparative Polyacrylamide Gel Electrophoresis and electroelution. The method is based on the observation that the high-molecular-weight buffer/SDS-soluble mucins do not penetrate through the Polyacrylamide Gel, but remain on the Gel surface. Mucus solution extracted with 6 M urea was fractionated on Sepharose CL-2B column and V0 peak mucin was submitted to purification by preparative Polyacrylamide Gel Electrophoresis (22 h). Nonpenetrated mucin layer was electroeluted from the Gel after the reversing of electrode polarity (3 h). A comparison of mucin preparations purified by our method and by CsC1 density gradient centrifugation indicated that the GalNAc/protein and GalNAc/DNA ratios were three times higher than those of the first method. The method is a relatively short and efficient procedure and yields pure mucin preparation free of contaminating proteins and nucleic acids.
Francisco E. Nicolás - One of the best experts on this subject based on the ideXlab platform.
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Purification of DNA Oligos by denaturing Polyacrylamide Gel Electrophoresis (PAGE).
Methods in enzymology, 2013Co-Authors: Sara Lopez-gomollon, Francisco E. NicolásAbstract:After chemical synthesis, the oligonucleotide preparation contains the desired full-length oligonucleotide but also all of the DNA molecules that were aborted during each cycle in the synthesis, and the by-products generated during the chemical reactions. The purification of oligonucleotides is a critical step for demanding applications where the exact length or sequence of the oligonucleotide is important, or for oligonucleotides longer than 50 bases. There are several methods of increasing oligonucleotide purity, the choice of which will depend on modifications of the oligonucleotides and their intended use. Polyacrylamide Gel purification (PAGE purification) is the method of choice when the highest percentage of full-length oligonucleotide is desired. This chapter describes a protocol for oligonucleotide purification using denaturing Polyacrylamide Gel Electrophoresis, and includes oligonucleotide preparation, Polyacrylamide Gel Electrophoresis, and purification from the Gel slice by two different methods: by diffusion or by electroelution. This chapter also includes recommendations as well as protocol advice.
Alexey Petrov - One of the best experts on this subject based on the ideXlab platform.
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RNA purification by preparative Polyacrylamide Gel Electrophoresis.
Methods in enzymology, 2013Co-Authors: Alexey Petrov, Elisabetta Viani Puglisi, Joseph D. PuglisiAbstract:Preparative Polyacrylamide Gel Electrophoresis (PAGE) is a powerful tool for purifying RNA samples. Denaturing PAGE allows separation of nucleic acids that differ by a single nucleotide in length. It is commonly used to separate and purify RNA species after in vitro transcription, to purify naturally occurring RNA variants such as tRNAs, to remove degradation products, and to purify labeled RNA species. To preserve RNA integrity following purification, RNA is usually visualized by UV shadowing or stained with ethidium bromide or SYBR green dyes.