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41.
In this study we describe the activation with chloroformates of Trisacryl-GF-2000, a new synthetic gel support that is stable,
hydrophilic, and contains large amounts of hydroxyl groups available for activation.
Of all the reagents tested, the activation withN-hydroxysuccinimide-chloroformate andp-nitrophenylchloroformate in organic solvents provides the best activation yield and subsequent coupling. When Trisacryl was
activated in acetone with the chloroformates in the presence of 4-dimethylaminopyridine as base and catalyst, up to 30% of
the hydroxyl groups, (i.e., 1/repeating unit) could be activated. Amino-containing ligands and proteins could be coupled to
these carriers at pH 8 or higher. For better results in affinitychromatographic applications, spacers of ε-amino caproic acid
or diaminohexane were introduced. The efficacy of these columns was demonstrated by purification of enzymes, antibodies, and
antigens. The performance of these new columns were compared with that of Sepharose columns activated in various ways. In
every case, the properties of the Trisacryl support proved superior with particular reference to the purity of the product
obtained. 相似文献
42.
Vlatakis G. Skarpelis G. Stratidaki I. Bouriotis V. Clonis Y. D. 《Applied biochemistry and biotechnology》1987,15(3):201-212
The resolution of restriction endonucleases from the same microorganism is conventionally achieved by lengthy fractionation
protocols. We now report effective single-step procedures that exploit dye-ligand chromatography for the resolution and purification
of restriction enzymes. After suitable initial screening, we demonstrated that resolution of two restriction activites can
be achieved in one chromatographic step, and further purification can subsequently be effected using selected dye-adsorbents.
Accordingly, we resolved in one step, Hpa I from Hpa II, Hind II from Hind III, and Sac I from Sac II. Furthermore, a three-step
Chromatographic procedure has been developed to purify EcoRV suitable for commercial exploitation, as judged by the “overdigestion”
and “cut-ligate-recut” quality control tests. 相似文献
43.
阴阳离子型表面活性剂双水相萃取色氨酸衍生物和牛血清白蛋白 总被引:7,自引:0,他引:7
溴化十二烷基三乙胺(C12NE)和十二烷基硫酸钠(SDS)在一定条件下混合可以形成具有清晰界面的两个水相,称为阴阳离子型表面活性剂双水相。作者研究了利用该双水相萃取3种色氨酸衍生物和牛血清白蛋白的可能性.文中采用工作曲线校正扣除表面活性剂背景的影响,萃取结果准确可信。 相似文献
44.
45.
The linear finite difference Poisson-Boltzmann (FDPB) equation is applied to the calculation of the electrostatic binding free energies of a group of inhibitors to the Neuraminidase enzyme. An ensemble of enzyme-inhibitor complex conformations was generated using Monte Carlo simulations and the electrostatic binding free energies of subtly different configurations of the enzyme-inhibitor complexes were calculated. It was seen that the binding free energies calculated using FDPB depend strongly on the configuration of the complex taken from the ensemble. This configurational dependence was investigated in detail in the electrostatic hydration free energies of the inhibitors. Differences in hydration energies of up to 7 kcal mol–1 were obtained for root mean square (RMS) structural deviations of only 0.5 Å. To verify the result, the grid size and parameter dependence of the calculated hydration free energies were systematically investigated. This showed that the absolute hydration free energies calculated using the FDPB equation were very sensitive to the values of key parameters, but that the configurational dependence of the free energies was independent of the parameters chosen. Thus just as molecular mechanics energies are very sensitive to configuration, and single-structure values are not typically used to score binding free energies, single FDPB energies should be treated with the same caution. 相似文献
46.
The growth ofBacillus amyloliquefaciens in the aqueous two-phase system, made up of polyethylene glycol, dextran, and water, was investigated. Generally,Bacillus partitions in the dextran phase, but the magnitude of the separation depends largely on the overall composition of polymers
in the phase system. The kinetics of growth ofBacillus amyloliquefaciens was studied in the polyethylene glycol-rich continuous phase, dextran-rich dispersed phase, and in the mixed phase. From
the kinetic data it appears that increasing the overall polymer composition causes the cells to adsorp at the interface. On
the other hand, partition measurements indicate that increasing polymer concentrations make the cell partitioning more one-sided.
This anomaly is explained by studying the interfacial adsorption of cells via dynamic surface tension measurements. 相似文献
47.
Remedios González-Luque Manuela Merchán Piotr Borowski Björn O. Roos 《Theoretical chemistry accounts》1993,86(6):467-476
Summary Multiconfigurational electron correlation methods have been analyzed in order to theoretically compute the electron affinity (EA) of ozone. The near-degeneracy correlation effects, which are so important in O3 and O
3
–
, have been described using complete active space (CAS) SCF wave functions. Remaining dynamic correlation effects are computed using second-order perturbation theory (the CASPT2 method). The best calculated adiabatic value (including zero-point energy corrections), 2.19 eV, is about 0.09 eV larger than the experimental value. Comparative studies using size-consistent coupled pair functional approaches (CPF and ACPF) have also been performed. The harmonic frequencies in O
3
–
have been determined to be: 1=992, 2=572, and 3=879 cm–1, which gives a zero-point energy of 0.151 eV. 相似文献
48.
亚氨基二乙酸—硅胶键合相的合成及其在核苷酸分析中的应用 总被引:2,自引:1,他引:2
以大孔微球硅胶(30nm,8μm)为基体,经γ-氨丙基三乙氧基硅烷活化与间隔臂氯代环氧丙烷反应后,再与螯合剂亚氨基二乙酸键合,后者与铜离子(Ⅱ)螯合后,构成定位金属离子亲和色谱固定相。用磷酸盐缓冲溶液(pH=7.0)作流动相,经紫外吸收检测(260nm)对核苷酸混合物进行了高效液相色谱分析,取得较满意的结果。 相似文献
49.
S. Bhattacharyya A. N. Sil T. K. Mukherjee P. K. Mukherjee P. Vasu 《International journal of quantum chemistry》2007,107(4):946-951
Electron affinity of plasma embedded muonium (μ+e?) and pionium (π+e?) has been estimated using a general three‐body formalism within variational framework. Electron correlation is taken care of using extended Hylleraas basis sets. The stability of the three‐body systems have been studied under the Debye screening model of the plasma, which changes the Coulombic potential to the screened Coulomb one. The electron affinity of the muonium and pionium is maximum for free systems, and it decreases gradually toward zero with increase of the plasma strength, which tends to push the three‐body system toward gradual instability. The electron correlation contribution to the affinity has been analyzed. © 2006 Wiley Periodicals, Inc. Int J Quantum Chem, 2007 相似文献
50.
Eric D BalighianJoel F Liebman 《Journal of fluorine chemistry》2002,116(1):35-39
It is well established that atomic fluorine has a miserly high ionization energy and avariciously high electron affinity. Yet, as was shown some 30 years ago, the latter value is anomalously low-linear extrapolation shows the value to be some 110 kJ/mol less than expected by comparison to the heavier halogens. Related literature investigation extended this anomaly to nitrogen and oxygen and their (valence) isoelectronic congeners. We extend it to related cationic species and thereby the noble gases and alkali metals, and the aforementioned species are better understood. We thus ask if the ionization energy/electron affinity anomaly of fluorine remains an anomaly. 相似文献