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11.
In CZE, the adsorption of the proteins on the capillary wall is a common problem. This paper describes the simple method of utilizing zwitterionic buffer additives to improve the separation of proteins in untreated fused silica capillaries at neutral pH. Three kinds of zwitterion are evaluated in the separation of acidic, neutral, and basic proteins, including their effect on protein efficiency, mobility, separation, and resolution; the difference between the effects of the different additives are also highlighted. The method has proved to be a possible means of reducing protein adsorption, especially for basic proteins. 相似文献
12.
13.
遗传算法是一种模仿自然进化进程的新颖的启发式优化方法。通过它,人们可以在计算机上对所需的各种问题实施进化操作,最终产生理想的结果。遗传算法现已被广泛于计算机辅助设计、工程设计、系统模拟等领域并取得了极大的成功。本文拟对遗传算法在对大、中、小分子的构象搜寻中的应用作一较全面的综述。 相似文献
14.
细胞色素b5Glu44,Glu56的定点突变和蛋白质结构比较 总被引:2,自引:0,他引:2
用寡聚核苷酸定点突变的方法将牛肝细胞色素b5基因上第44位和56位谷氨酸的密码子GAA变成氨酸的密码子GCT,获得突变体E44A,E56A和E44/56A的基因。 它们分别克隆于PUC19上,转化大肠杆菌JM83后,突变基因得到成功的表达。 相似文献
15.
A. W. Flounders D. L. Brandon A. H. Bates 《Applied biochemistry and biotechnology》1995,50(3):265-284
Monoclonal antibodies specific for thiabendazole were immobilized to silicon, silicon dioxide, stoichiometric silicon nitride,
and silicon-rich silicon nitride surfaces. This work provides the foundation for the development of a homogeneous sensor system
for rapid detection and quantification of thiabendazole residues in produce and animal tissue. Immobilization was performed
via aqueous silanization of the substrate followed sequentially by treatment with glutaraldehyde and contact with antibody
solution in the presence of detergent. Surfaces were challenged with thiabendazole-horseradish peroxidase conjugate in an
ELISA format to estimate immobilized antibody load. A stable and reproducible surface loading of 2 x 1011 antibodies/cm2 was obtained only after surfaces received postimmobilization treatments to remove nonspecifically adsorbed antibody. No difference
in surface loading was noted when using 30% hydrogen peroxide rather than nitric acid for silanol activation. Little difference
was noted among the antibody loadings achieved on the various silicon substrates. Bound antigen-enzyme conjugate was eluted
with 0.1N acetic acid and reproducible surface activity was measured for up to four consecutive antigen challenges. Immobilized
antibody surfaces were stabilized with 2% sucrose, dehydrated at 37‡C and stored in vacuum or stored at 4‡C in phosphate buffered
saline containing 0.01% sodium azide without significant loss of activity. 相似文献
16.
The monodisperse, porous poly(chloromethylstyrene-co-divinylbenzene) beads of 7.9 microm were prepared by a single-step swelling and polymerization method. The seed particles prepared by dispersion polymerization exhibited good absorption of the monomer phase. Based on this media, a weak cation-exchange (WCX) stationary phase for HPLC was synthesized by a new chemically modified method. The prepared resin has advantages of biopolymer separation, high column efficiency, low column backpressure, high protein mass recovery, and good resolution for proteins. The dynamic protein-loading capacity of the synthesized WCX packings was 18.2 mg/g. Five proteins were separated in 3.0 min using the synthesized WCX stationary phase. The experimental results show that the obtained WCX resin has very weak hydrophobicity. The WCX resin was also used for the rapid separation and purification of lysozyme from egg white in 5.0 min with only one step. The purity and specific bioactivity of the purified lysozyme were found to be more than 93% and 70 245 U/mg, respectively. 相似文献
17.
A.G. Dobrikova M.I. Dimitrov S.G. Taneva I.B. Petkanchin 《Colloids and surfaces. B, Biointerfaces》2007,56(1-2):114-Optics
Using microelectrophoresis and electric light scattering techniques, we investigated the adsorption characteristics, surface coverage and surface electric parameters of superstructures from two isoforms of plastocyanin, PCa and PCb, in an oxidized state adsorbed on β-ferric hydrous oxide particles. The surface electric charge and electric dipole moments of the composite particles and the thickness of the protein adsorption layer are determined in a wide pH range, at different ionic strengths and concentration ratios of PC to β-FeOOH. The adsorption of the two proteins was found to shift the particles’ isoelectric point and to alter the total electric charge and the electric dipole moments of the oxide particles to different extent. A “reversal” in the direction of the permanent dipole moment is observed at lower pH for PCb- than for PCa-coated oxide particles. Strict correlation is found between the changes in the electrokinetic charge of the composite particles and the variation in their “permanent” dipole moments. Data suggest that the adsorption of the proteins is driven by electrostatic and/or hydrophobic interactions with the oxide surfaces dependent on pH. The adsorption behaviour is consistent with the involvement of the “eastern” and “northern” patches of the plastocyanin molecules in their adsorption on the oxide surfaces that are differently charged depending on pH. 相似文献
18.
脑神经退行性疾病中的有机化学-朊病毒(疯牛病)中的蛋白物理有机化学和自由基化学 总被引:5,自引:0,他引:5
杨池明 《高等学校化学学报》2002,23(2):243-250
通过关于“普里昂”蛋白病毒疾病的已有临床、医学生理、免疫和化学等方面的现象,讨论了朊病毒当中的部分蛋白氧化损伤和蛋白自由基化学本质。 相似文献
19.
The ability to recover and purify natural and recombinant proteins, and the costs of doing so remain a major task in introducing
the potential products of biotechnology. The bases for separation range from specific binding onto tailored reagents to solubility
and partitioning behavior governed by a mixed bag of size, charge, and hydrophobicity. In most cases, a combination of methods
is used in sequence, and improvements in the selectivity at an early stage can enhance the effectiveness of subsequent (and
usually more costly) steps. Genetic engineering provides a means of improving the selectivity within the context of existing
separation methods.
By this strategy, improvements in selectivity are sought by bestowing a distinctive property on the protein of interest. The
primary sequence of amino acids is altered, such that the protein can be selectively removed from other components of the
multicomponent mixture in which such products are commonly found. In this article, the range of these “distinctive properties”
and their pairing with various separation methods will be reviewed. Specific examples from our work, in which a distinctive
charge is provided via a polypeptide “purification” fusion tail, will be discussed. Separation methods we have used with these
fusion proteins are precipitation, two-phase aqueous extraction, reversed micellar extraction, and ion exchange using both
resins and membranes. 相似文献
20.
In this paper, the spectral behavior of protein and Poniacyl Carmine 2B (PC 2B) has been studied by spectrophotometric method. The conditional constants, apparent combination constant K and maximum binding number n, were used to express the combination ability of the reactions between PC 2B and protein under a set of given conditions. The Sandell index s was used to express the sensitivity of the determination of protein. The factors, acidity, PC 2B concentration and the ionic strength, were discussed by the change of apparent combination constant and maximum binding number. It was found that acidity of the solution, PC 2B concentration and ionic strength had a significant effect on the sensitivity of the assay of protein. Under the optimal conditions, the apparent combination constant K and the maximum binding number n were 2.36 × 106 L mol?1 and 95, respectively. With further investigation, it was found that the Scatchard model was suitable in treating the data obtained in the experiments. In the buffer medium of HCl‐KCl at 1.87, the addition of protein made the maximum absorption of the system move from 527 nm to 513 nm. Its apparent molar absorptivity is 4.46 × 105 Lmol?1 cm?1 at 513 nm. Beer's law is obeyed in the range of 0 ? 55 μg mL?1. The system developed in this paper has been used for the determination of protein in milk powder successfully. 相似文献