Chemically modified porous silica gel as a bioadsorbent and a biocatalyst.
TL;DR: Epoxy silica gel, prepared by reaction of porous silica Gel with γ-glycidoxypropyltriethoxysilane, was coupled with m -aminobenzamidine and the resulting matrix was used for trypsin purification.
About: This article is published in Analytical Biochemistry.The article was published on 1979-09-15. It has received 12 citations till now. The article focuses on the topics: Silica gel & Human serum albumin.
Citations
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TL;DR: It was found that treatment of gels with dithiothreitol prior to impregnation with silver nitrate results in more reproducible staining patterns that are also qualitatively similar to those obtained with Coomassie blue.
4,022 citations
TL;DR: The AMPPD technique was demonstrated by the detection of proteins from individual photoreceptor outer segments, including proteins constituting approximately 1% of the total, and proved extremely sensitive for immunoblotting.
Abstract: The analysis of proteins from single cells requires techniques of supreme sensitivity. Although radiochemical procedures are capable of detecting small amounts of electrophoretically separated proteins, their sensitivity falls short of that required for routine detection of minor components of single cells. Utilizing the avidin-biotin interaction and the alkaline phosphatase substrate 3-(4-methoxyspiro[1,2-dioxetane-3,2'- tricyclo-[3.3.1.1(3,7)]decan]-4-yl)phenyl phosphate (AMPPD), we have developed an alternative, chemiluminescence-based method for protein detection whose sensitivity exceeds that of other methods. Applying this method to a purified protein, we could detect as little as 63 fg (0.9 amol) of biotinylated bovine serum albumin. The sensitivity of the method was demonstrated by the detection of proteins from individual photoreceptor outer segments, including proteins constituting approximately 1% of the total. Chemiluminescence detection also proved extremely sensitive for immunoblotting: a comparison of five methods for detection of antibody-antigen interactions showed that the AMPPD technique was more sensitive than detection with a colorimetric alkaline phosphatase substrate, 125I-labeled protein A, 125I-labeled anti-mouse IgG, or colloidal gold-conjugated anti-mouse IgG.
93 citations
TL;DR: The first two non-silanizing coupling methods are simple, inexpensive and non-hazardous compared to the third, more complex method in which an initial Correspondance: to PVS.
Abstract: Several methods for immobilizing anti-T2 mycotoxin monoclonal antibodies on quartz fibers, for use in optical sensor development, have been evaluated with respect to the surface density and stability of the immobilized proteins. the first method activates matrix hydroxyl groups using p-toluenesulfonyl chloride (TSC). the second method activates these groups using p-nitrophenyl chloroformate (NPCF). the third method requires an initial silanization using 3-aminopropyltriethoxysilane (APTES) followed by carrier activation with glutaraldehyde. the activated carrier in all three methods is then reacted with the amino groups of the protein. the first two non-silanizing coupling methods are simple, inexpensive and non-hazardous compared to the third, more complex method in which an initial Correspondance: to PVS
39 citations
TL;DR: A rapid (6.5 min) and simple one-step magnetic immunoassay (MIA) has been developed for analysis of human urinary albumin in near patient settings and the obtained results showed good correlation with the hospital immunoturbidimetric reference method.
29 citations
TL;DR: The environmental parameters necessary for increased production of mature, 35‐kDa active protease in strains of L. monocytogenes are described and its detection using polyclonal antibodies raised against Bacillus subtilis neutral protease is described.
Abstract: Listeria monocytogenes, a facultative intracellular pathogen, synthesizes an extracellular protease which is responsible for the maturation of phosphatidylcholine phospholipase C (lecithinase), a virulence factor involved in cell-to-cell spread. This work describes the environmental parameters necessary for increased production of mature, 35-kDa active protease in strains of L. monocytogenes, and its detection using polyclonal antibodies raised against Bacillus subtilis neutral protease. High performance liquid affinity chromatography was exploited to isolate the biologically active form of the mature protease, which was then subjected to biochemical characterization using casein as a substrate. The protease is a zinc-dependent metalloprotease which degrades casein over a wide range of temperatures and pH values. It can also degrade actin, the most abundant protein in many eukaryotic cells. The Listeria protease was shown to exhibit a high thermal stability and a relatively narrow substrate specificity. A three-dimensional model built on the basis of the homology with thermolysin was used to understand the structural basis of these characteristics.
26 citations
Cites methods from "Chemically modified porous silica g..."
...Prior to coupling of bacitracin to the silica, the latter was activated by the addition of an epoxy group (Roy and Kundu 1979; van den Burg et al. 1989)....
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References
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TL;DR: Anti-globoside is directed toward the terminal β-(N-acetyl)galactosaminosyl-(1→3)α-galactopyranosyl structure, while anti-hematosides is directed predominantly toward the sialosyl residue of hematoside.
59 citations
TL;DR: DEAE-silica gel has been shown to be an improvement over DEAE-Sephadex for the quantitative isolation of gangliosides and neutral glycosphingolipids from animal tissues or cells and preliminary results indicated that it can also be used for protein separation.
41 citations
TL;DR: In this paper, the deviations from the stoichiometric composition of HgTe and Hg 0.82 Cd 0.18 Te crystals have been controlled by heat treatment under Hg vapor pressure.
35 citations
TL;DR: In this article, the effect of salt concentration and pH of eluent on protein elution volumes from aminosilochrom columns is discussed, and it is possible to use the aminosILochrom for chromatographic separation of proteins.
Abstract: Adsorption and chromatography of several proteins on macroporous silica — Silochrom was investigated. Chemical modification of the Silochrom surface by means of γ-aminopropyltriethoxysilane eliminates irreversible protein adsorption. The effect of salt concentration and pH of eluent on protein elution volumes from “aminosilochrom” columns is discussed. It is possible to use the aminosilochrom for chromatographic separation of proteins.
34 citations
TL;DR: Results indicated that DEAE-silica gel is preferable to the other two ion exchangers for the fractionation of ganglioside mixtures.
31 citations