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101.
Preparation of a novel lysozyme molecularly imprinted polymer using uniformly sized functionalized poly(glycidyl methacrylate) microspheres as the matrix and its application to lysozyme purification 下载免费PDF全文
Jing Zhang Yong‐le Tian Ling‐ling Wang Yan‐ting Han 《Biomedical chromatography : BMC》2014,28(4):534-540
A novel lysozyme imprinted polymer based on uniformly sized functionalized poly(glycidyl methacrylate) microspheres has been synthesized in aqueous solution using the surface imprinting technique. The microspheres were modified with hydroxyl ethyl methacrylate to allow for the introduction of polymerizable double bonds, with β‐cyclodextrin and acrylamide being grafted onto the surface as functional monomers. The selective recognition properties of the resulting molecularly imprinted polymers (MIPs) were investigated by HPLC. Various factors were also investigated in terms of their influence on the retention behaviors of the imprinted polymers, including the pH and salt concentration of the mobile phase. The binding capability properties of the MIPs were evaluated, and the PGMA/EDMA‐MIPs showed a high adsorption capacity for lysozyme. Furthermore, this MIP was used to separate and enrich lysozyme from egg whites. The results revealed that the lysozyme surface‐modified MIP could be used to efficiently separate and purify lysozyme from egg whites. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
102.
Based on three-state renaturation process of denatured proteins, an equation describing the effect of denaturant concentration on renaturation yield of denatured proteins was presented. By this equation, two parameters n(m1 -m2) and Ka can be obtained. The former indicates the difference in the number of denaturant molecules between the renaturation process of n number of refolding intermediates from refolding intermediate state to native state and their aggregate process from refolding intermediate state to aggregate state, the latter denotes the apparent aggregate equilibrium constant for protein molecules aggregated from native state to aggregate state, and from them, the characteristics of the renaturation process of denatured proteins in denaturant solution can be identified. This equation was tested by the renaturation processes of denatured egg white lysozyme in guanidine hydrochloride and urea solutions, with the results to show that when guanidine hydrochloride and urea concentrations were separately higher than 1.25 and 3.00 mol/L or separately lower than 1.00 and 3.00 mol/L, the refolding intermediates of egg white lysozymes were more easily aggregated to aggregate state or more easily renatured to native state, respectively. Under different initial total egg white lysozyme concentrations in urea solution, the refolding egg white lysozyme intermediates could be deduced to have a tendency to form a bimolecular intermediate aggregate, and this inference was further confirmed by their nonreducing SDS-PAGE and size exclusion chromatography. 相似文献
103.
The interactions between seven anionic azo dyes and hen egg-white lysozyme were investigated by means of visible absorption spectroscopic and lytic activity measurements. The dyes, for which no report on the interaction has appeared so far, were bound with lysozyme to different extents as a result of the differences in their chemical structures. Although most of the dyes formed 11 complex with lysozyme, two of them behaved differently. From the measurements of the absorption, lytic inhibition toward cell walls and competitive binding with substrate analogues, it was concluded that a major binding site of these dyes on lysozyme is a charged lysine residue in the vicinity of the subsitesD, E andF, and, in the case of a dye that forms 21 complex with lysozyme, and additional binding site is in the vicinity of the subsite B. 相似文献
104.
105.
Polarographic Catalytic Wave of Lysozyme 总被引:2,自引:0,他引:2
Lysozyme(LE)isatypicalproteinwithwell-knownstrUcture.Itcontainsfourdisulfidebonds,andthreeofthemarewraPpedinsideLEmolecule.Thedisulfidebondbetweencys6andcysl27isexposedexternallyinthehydrophilicregi0n.ItwasfoundthatLEyieldedapolarograPhicwaveinO.2mollLNaAc~HAc(pH4.7)buffersolution.Therewereareductionwave(IP,c)andanoxidationwave(IP,a)inthecyclicvoltanunogram.ThepeakcurrentratiobetweenIP,aandIP,cwasl:l.ThepeakpotentialEP,cofthereductionwavewas-().53V(vs,SCE),andtheEP,aoftheoxidatio… 相似文献
106.
The apparent molar volumes, V,2, of glycine, alanine, -amino-n-butyric acid, valine, leucine, and lysine monohydrochloride have been determined in aqueous solutions of 0.05, 0.1, and 0.4 mol-kg–1 Triton X-100 (TX-100), and the partial specific volume, v0, of hen-egg-white lysozyme in 0.4 mol-kg–1 TX-100 by density measurements at 298.15 K. These data have been used to calculate the infinite dilution apparent molar volumes, V2,m0, for the amino acids in aqueous TX-100 solutions and the standard partial molar volumes of transfer, tr V2,m0, of the amino acids from water to the aqueous surfactant solutions. The linear correlation of V2,m0 for a homologous series of amino acids has been utilized to calculate the contribution of the charged end groups (NH3+, COO–), CH2 group and other alkyl chains of the amino acids to V2,m0. The results on tr V2,m0, of amino acids from water to aqueous TX-100 solutions have been interpreted in terms of ion–ion, ion–polar, hydrophilic–hydrophilic and hydrophobic–hydrophobic group interactions. For all the six amino acids studied, the values of tr V2,m0 from water to all the studied concentrations of aqueous TX-100 are small in spite of their different hydrophobic content, indicating an overall balance in interactions of zwitterionic/hydrophilic groups of amino acids with the hydrophilic groups of TX-100, and of hydrophobic and ionic/hydrophilic groups of the amino acids with hydrophobic groups of TX-100. Comparison of the interactions of the amino acids with nonionic, anionic and cationic surfactants has also been made and discussed. The partial specific volume of transfer of lysozyme from water to aqueous TX-100 solutions also indicates a balance of the hydrophobic and hydrophilic interactions in the protein–nonionic surfactant system. 相似文献
107.
用动态光散射法研究了不同浓度NaCl对液—液扩散法生长溶菌酶晶体的影响, 并测量了晶体生长前后体系的Zeta电势.结果表明,NaCl浓度较高时,在溶菌酶溶 液—凝胶界面处会发生液液分层现象,溶液中一直存在较大的聚集体,生长出的晶 体质量较差.而在合适的NaCl浓度下,随着溶液Zeta电势降低,溶液中溶菌酶的大 的聚集体发生解聚集,生长出的晶体质量较高. 相似文献
108.
YanWANG BoLinGONG XinDuGENG 《中国化学快报》2003,14(8):828-831
Oxidative refolding of the denatured/reduced lysozyme was investigated by using weak-cation exchange chromatography (WCX). The stationary phase of WCX binds to the reduced lysozyme and prevented it from forming intermolecular aggregates. At the same time urea and ammonium sulfate were added to the mobile phase to increase the elution strength for lysozyme. Ammonium sulfate can more stabilize the native protein than a common eluting agent,sodium chloride. Refolding of lysozyme by using this WCX is successfully. It was simply carried out to obtain a completely and correctly refolding of the denatured lysozyme at high concentration of 20.0 mg/mL. 相似文献
109.
X. P. Geng M. R. Zheng B. H. Wang Z. M. Lei X. D. Geng 《Journal of Thermal Analysis and Calorimetry》2008,93(2):503-508
Calorimetric measurement of adsorption enthalpies of native lysozyme(Lyz) on a moderately hydrophobic surface at 25°C, pH
7.0 and various salt concentrations was performed. Based on the thermodynamics of stoichiometric displacement theory (SDT),
we calculated the fractions of thermodynamic functions involving four subprocesses during a displacement adsorption process
from the directly determined enthalpies in combination with adsorption isotherm measurements. The thermodynamic fractions
reveal the relative degree of the four subprocesses for contributions to enthalpy, entropy and free energy. The results show
that native Lyz adsorption on a moderately hydrophobic surface is an entropy driven process contributed mainly by conformational
loss of adsorbed Lyz. 相似文献
110.
Approaches for improving the performance of near-infrared diffuse reflectance spectroscopy (NIRDRS) have been extensively studied. The silver mirror was found to be a substrate to enhance the detection ability of this technique. For further optimization of the method, experimental and chemometric efforts were made in this study. Lysozyme samples of different concentrations were spotted on the silver mirror for measuring the spectra. Low-resolution spectra with repeated measurements were used for increasing the signal-to-noise ratio, and the chemometric methods of variable selection and nonlinear modeling were used to make the quantitative model more effective and accurate. The results show that the quality of the spectra was improved and a satisfactory quantitative model was obtained after optimization. The maximum deviation for the prediction samples is only 2.98?µg, which is much lower than the reported values in previous papers. Significant advances were obtained for NIRDRS for trace analysis. 相似文献