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51.
Effects of purification on the crystallization of lysozyme 总被引:2,自引:0,他引:2
Felecia L. Ewing Elizabeth L. Forsythe Mark van der Woerd Marc L. Pusey 《Journal of Crystal Growth》1996,160(3-4):389-397
We have additionally purified a commercial lysozyme preparation by cation exchange chromatography, followed by recrystallization. This material is 99.96% pure with respect to macromolecular impurities. At basic pH, the purified lysozyme gave only tetragonal crystals at 20°C. Protein used directly from the bottle, prepared by dialysis against distilled water, or which did not bind to the cation exchange column had considerably altered crystallization behavior. Lysozyme which did not bind to the cation exchange column was subsequently purified by size exclusion chromatography. This material gave predominately bundles of rod-shaped crystals with some small tetragonal crystals at lower pHs. The origin of the bundled rod habit was postulated to be a thermally dependent tetragonal ↔ orthorhombic change in the protein structure. This was subsequently ruled out on the basis of crystallization behavior and growth rate experiments. This suggests that heterogeneous forms of lysozyme may be responsible. These results demonstrate three classes of impurities: (1) small molecules, which may be removed by dialysis; (2) macromolecules, which are removable by chromatographic techniques; and (3) heterogeneous forms of the protein, which can be removed in this case by cation exchange chromatography. Of these, heterogeneous forms of the lysozyme apparently have the greatest affect on its crystallization behavior. 相似文献
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Tho Duc Khanh Nguyen Stefania Rabasco Alicia A. Lork Andre Du Toit Prof. Andrew G. Ewing 《Angewandte Chemie (International ed. in English)》2023,62(28):e202304098
We used correlative transmission electron microscopy (TEM) and nanoscale secondary ion mass spectrometry (NanoSIMS) imaging to quantify the contents of subvesicular compartments, and to measure the partial release fraction of 13C-dopamine in cellular nanovesicles as a function of size. Three modes of exocytosis comprise full release, kiss-and-run, and partial release. The latter has been subject to scientific debate, despite a growing amount of supporting literature. We tailored culturing procedures to alter vesicle size and definitively show no size correlation with the fraction of partial release. In NanoSIMS images, vesicle content was indicated by the presence of isotopic dopamine, while vesicles which underwent partial release were identified by the presence of an 127I-labelled drug, to which they were exposed during exocytosis allowing entry into the open vesicle prior to its closing again. Demonstration of similar partial release fractions indicates that this mode of exocytosis is predominant across a wide range of vesicle sizes. 相似文献
54.
Richard Ewing Raytcho Lazarov Yanping Lin 《Numerical Methods for Partial Differential Equations》2000,16(3):285-311
In this article, we study finite volume element approximations for two‐dimensional parabolic integro‐differential equations, arising in the modeling of nonlocal reactive flows in porous media. These types of flows are also called NonFickian flows and exhibit mixing length growth. For simplicity, we consider only linear finite volume element methods, although higher‐order volume elements can be considered as well under this framework. It is proved that the finite volume element approximations derived are convergent with optimal order in H1‐ and L2‐norm and are superconvergent in a discrete H1‐norm. By examining the relationship between finite volume element and finite element approximations, we prove convergence in L∞‐ and W1,∞‐norms. These results are also new for finite volume element methods for elliptic and parabolic equations. © 2000 John Wiley & Sons, Inc. Numer Methods Partial Differential Eq 16: 285–311, 2000 相似文献
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David F. Ewing Christophe Len Grahame Mackenzie Gino Ronco Pierre Villa 《Tetrahedron: Asymmetry》2000,11(24)
1,2-O-Isopropylidene-α-
-xylofuranose has been used to protect one aldehyde group of o-phthalaldehyde. This chiral protecting group acts as a resolving agent and this leads to separable diastereoisomers when a new stereogenic centre is created by the conversion of the second aldehyde group to a benzyloxyhydroxyethyl chain. These separated diastereoisomers were cyclised to 1,3-dihydrobenzo[c]furans with retention of chiral integrity at the C3 site thus allowing further elaboration to enantiomerically pure nucleoside analogues. 相似文献
57.
Treatment of a primary t-butyl carbamate (Boc) group with excess sodium t-butoxide in slightly wet tetrahydrofuran or 2-methyltetrahydrofuran provides the corresponding primary amine in excellent yield. We believe the reaction proceeds through an isocyanate intermediate. 相似文献
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The preparation and characterization of new tetrahalodimolybdenum(II) complexes with alkyl-substituted pyridines is described. IR and visible data are used to infer structural information with respect to previously prepared complexes of this type. 相似文献