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Open accessJournalISSN: 0021-9258

Journal of Biological Chemistry

About: Journal of Biological Chemistry is an academic journal. The journal publishes majorly in the area(s): Phosphorylation & Signal transduction. It has an ISSN identifier of 0021-9258. It is also open access. Over the lifetime, 193509 publication(s) have been published receiving 15784928 citation(s). The journal is also known as: JBC & J. Biol. Chem.. more


Open accessJournal Article
Abstract: Since 1922 when Wu proposed the use of the Folin phenol reagent for the measurement of proteins, a number of modified analytical procedures utilizing this reagent have been reported for the determination of proteins in serum, in antigen-antibody precipitates, and in insulin. Although the reagent would seem to be recommended by its great sensitivity and the simplicity of procedure possible with its use, it has not found great favor for general biochemical purposes. In the belief that this reagent, nevertheless, has considerable merit for certain application, but that its peculiarities and limitations need to be understood for its fullest exploitation, it has been studied with regard to effects of variations in pH, time of reaction, and concentration of reactants, permissible levels of reagents commonly used in handling proteins, and interfering substances. Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein. more

Topics: Reagent (61%), Sulfhydryl reagent (61%), Folin–Ciocalteu reagent (61%) more

285,427 Citations

Open accessJournal ArticleDOI: 10.1016/S0021-9258(18)64849-5
Abstract: Work from this laboratory resulted in the development of a method for the preparation and purification of brain lipides (1) which involved two successive operations. In the first step, the lipides were extracted by homogenizing the tissue with 2: 1 chloroform-methanol (v/v), and filtering the homogenate. In the second step, the filtrate, which contained the tissue lipides accompanied by non-lipide substances, was freed from these substances by being placed in contact with at least 5-fold its volume of water. This water washing entailed the loss of about 1 per cent of the brain lipides. This paper describes a simplified version of the method and reports the results of a study of its application to different tissues, including the efficiency of the washing procedure in terms of the removal from tissue lipides of some non-lipide substances of special biochemical interest. It also reports some pertinent ancillary findings. The modifications introduced into the method pertain only to the washing procedure. A chloroformmethanol extract of the tissue, prepared as described in the original version of the method, is mixed with 0.2 its volume of water to which, for certain purposes, different mineral salts may be added. A biphasic system without any interfacial fluff is obtained (2). The upper phase contains all of the non-lipide substances, most of the strandin, and only negligible amounts of the other lipides. The lower phase contains essentially all the tissue lipides other than strandin. In comparison with the original method, the present version has the advantage of being simpler, of being applicable to any scale desired, of substantially decreasing the losses of lipides incidental to the washing process, and, finally, of yielding a washed extract which can be taken to dryness without foaming and without splitting of the proteolipides (3). more

Topics: Triglyceride transport (51%)

56,823 Citations

Open accessJournal ArticleDOI: 10.1016/S0021-9258(19)83641-4
Abstract: A new family of highly fluorescent indicators has been synthesized for biochemical studies of the physiological role of cytosolic free Ca2+. The compounds combine an 8-coordinate tetracarboxylate chelating site with stilbene chromophores. Incorporation of the ethylenic linkage of the stilbene into a heterocyclic ring enhances the quantum efficiency and photochemical stability of the fluorophore. Compared to their widely used predecessor, "quin2", the new dyes offer up to 30-fold brighter fluorescence, major changes in wavelength not just intensity upon Ca2+ binding, slightly lower affinities for Ca2+, slightly longer wavelengths of excitation, and considerably improved selectivity for Ca2+ over other divalent cations. These properties, particularly the wavelength sensitivity to Ca2+, should make these dyes the preferred fluorescent indicators for many intracellular applications, especially in single cells, adherent cell layers, or bulk tissues. more

Topics: Fluorophore (53%), Fluorescence (53%)

21,254 Citations

Open accessJournal ArticleDOI: 10.1016/S0021-9258(18)94333-4
Klaus Weber1, Mary Osborn1Institutions (1)
Abstract: Forty proteins with polypeptide chains of well characterized molecular weights have been studied by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate following the procedure of Shapiro, Vinuela, and Maizel (Biochem. Biophys. Res. Commun., 28, 815 (1967)). When the electrophoretic mobilities were plotted against the logarithm of the known polypeptide chain molecular weights, a smooth curve was obtained. The results show that the method can be used with great confidence to determine the molecular weights of polypeptide chains for a wide variety of proteins. more

19,356 Citations

Open accessJournal ArticleDOI: 10.1016/S0021-9258(19)41496-8
Abstract: A technique has been developed for the separation of proteins by two-dimensional polyacrylamide gel electrophoresis. Due to its resolution and sensitivity, this technique is a powerful tool for the analysis and detection of proteins from complex biological sources. Proteins are separated according to isoelectric point by isoelectric focusing in the first dimension, and according to molecular weight by sodium dodecyl sulfate electrophoresis in the second dimension. Since these two parameters are unrelated, it is possible to obtain an almost uniform distribution of protein spots across a two-diminsional gel. This technique has resolved 1100 different components from Escherichia coli and should be capable of resolving a maximum of 5000 proteins. A protein containing as little as one disintegration per min of either 14C or 35S can be detected by autoradiography. A protein which constitutes 10 minus 4 to 10 minus 5% of the total protein can be detected and quantified by autoradiography. The reproducibility of the separation is sufficient to permit each spot on one separation to be matched with a spot on a different separation. This technique provides a method for estimation (at the described sensitivities) of the number of proteins made by any biological system. This system can resolve proteins differing in a single charge and consequently can be used in the analysis of in vivo modifications resulting in a change in charge. Proteins whose charge is changed by missense mutations can be identified. A detailed description of the methods as well as the characteristics of this system are presented. more

18,529 Citations

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Journal's top 5 most impactful authors

P.A. Levene

346 papers, 3.7K citations

Robert D. Simoni

239 papers, 1.3K citations

Robert L. Hill

235 papers, 871 citations

C. A. Elvehjem

198 papers, 6.8K citations

Nicole Kresge

174 papers, 395 citations

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