Topic
Gel electrophoresis
About: Gel electrophoresis is a research topic. Over the lifetime, 26026 publications have been published within this topic receiving 1113565 citations.
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TL;DR: The purification and characterization of acetylcholinesterase from heads of the fruit fly Drosophila and the hydrolyzed butyrylthiocholine much more efficiently than these eel or human enzymes are described, an indication that the fly head enzyme has a substrate specificity intermediate between mammalian acetyl Cholinestersterases and butyRYlchlinesterases.
184 citations
TL;DR: Continuous electrophoresis buffers are described for polyacrylamide gels at pH values ranging from 3.8 to 10.2 and are likely to be useful in the detection of genetic variation in proteins and in other applications.
184 citations
TL;DR: The precise locations of the DNase I cutting sites in the nucleosome core have been determined by analysis of the DNA products of aDNase I digestion of 32P end-labelled mucleosome cores on a high resolution gel electrophoresis system and the stagger between individual sites on opposite strands has been determined and is found to be inconsistent with at least one proposed mechanism for nuclease cleavage of chromatin DNA.
Abstract: The precise locations of the DNase I cutting sites in the nucleosome core have been determined by analysis of the DNA products of a DNase I digestion of 32P end-labelled mucleosome cores on a high resolution gel electrophoresis system. This system is capable of resolving fragments of mixed sequence DNA differing by one base into the region of 160 bases in length. The DNase I cutting sites in the core are found to be spaced at multiples of about 10.4 (i.e. clearly different from 10.0) bases along the DNA, but show significant variations about this value. In addition to the location of the sites, the stagger between individual sites on opposite strands has been determined and is found to be inconsistent with at least one proposed mechanism for nuclease cleavage of chromatin DNA. Finally, a calculated distribution of fragment lengths in a DNase I digest of nuclei has been determined from the data obtained from the nucleosome core and found to be in reasonable agreement with the observed distribution. The periodicity of 10.4 is discussed with respect to the number of base pairs per turn of chromatin DNA and the number of superhelical turns of DNA per nucleosome.
184 citations
TL;DR: Carrier-protein harvesting provides a rich source of candidate peptides and proteins with potential diverse tissue and cellular origins that may reflect important disease-related information.
Abstract: Background: Albumin binds low–molecular-weight molecules, including proteins and peptides, which then acquire its longer half-life, thereby protecting the bound species from kidney clearance. We developed an experimental method to isolate albumin in its native state and to then identify [mass spectrometry (MS) sequencing] the corresponding bound low–molecular-weight molecules. We used this method to analyze pooled sera from a human disease study set (high-risk persons without cancer, n= 40; stage I ovarian cancer, n = 30; stage III ovarian cancer, n = 40) to demonstrate the feasibility of this approach as a discovery method.
Methods: Albumin was isolated by solid-phase affinity capture under native binding and washing conditions. Captured albumin-associated proteins and peptides were separated by gel electrophoresis and subjected to iterative MS sequencing by microcapillary reversed-phase tandem MS. Selected albumin-bound protein fragments were confirmed in human sera by Western blotting and immunocompetition.
Results: In total, 1208 individual protein sequences were predicted from all 3 pools. The predicted sequences were largely fragments derived from proteins with diverse biological functions. More than one third of these fragments were identified by multiple peptide sequences, and more than one half of the identified species were in vivo cleavage products of parent proteins. An estimated 700 serum peptides or proteins were predicted that had not been reported in previous serum databases. Several proteolytic fragments of larger molecules that may be cancer-related were confirmed immunologically in blood by Western blotting and peptide immunocompetition. BRCA2, a 390-kDa low-abundance nuclear protein linked to cancer susceptibility, was represented in sera as a series of specific fragments bound to albumin.
Conclusion: Carrier-protein harvesting provides a rich source of candidate peptides and proteins with potential diverse tissue and cellular origins that may reflect important disease-related information.
184 citations
TL;DR: In this paper, a novel separation method of metallic and semiconducting single-wall carbon nanotubes (SWCNTs) using agarose gel electrophoresis was developed.
Abstract: We have developed a novel separation method of metallic and semiconducting single-wall carbon nanotubes (SWCNTs) using agarose gel electrophoresis. When the SWCNTs were isolated with sodium dodecyl sulfate (SDS) and embedded in agarose gel, only the metallic SWCNTs separated from the starting gel by an electric field. After 20 min, almost all SWCNTs applied to gel electrophoresis were separated into two fractions, containing ~95% semiconducting and ~70% metallic nanotubes. The difference in the response to the electric field between metallic and semiconducting SWCNTs can be explained by the higher affinity of semiconducting SWCNTs to agarose than to SDS.
184 citations