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排序方式: 共有90条查询结果,搜索用时 31 毫秒
61.
建立了免疫亲和柱净化/高效液相色谱串联质谱法测定婴幼儿配方乳粉中生物素的方法。样品用磷酸盐缓冲液溶解,并通过生物素免疫亲和柱净化后,采用Ultimate AQ-C18(2.1 mm×100 mm,3μm)色谱柱分离,以甲醇-0.1%甲酸(30∶70)为流动相,进样量为5.0μL,流速为0.3 mL/min,柱温为30℃,并经电喷雾电离串联质谱在多离子反应监测(MRM)模式下进行测定,定量子离子为227.2,定性子离子为227.2、96.9,碰撞能量分别为10、30 V。结果显示,生物素在0.1~1.0μg/mL范围内线性关系良好,方法检出限为20.3μg/kg。对空白试样进行3个浓度水平的加标回收实验,测得加标回收率为92.7%~98.5%,相对标准偏差为1.7%~1.9%。该方法具有样品处理简单、灵敏度高、重现性好、分析时间短等优点,可以满足婴幼儿配方乳粉中生物素含量的测定要求。 相似文献
62.
63.
Immobilization of active facilitated glucose transporters (GLUT-1) in supported biological membranes
Ch. Neumann-Spallart F. Pittner Thomas Schalkhammer 《Applied biochemistry and biotechnology》1997,68(3):153-169
Membrane fragments or membrane proteins within a lipid mixture were immobilized over metal electrodes. This procedure has
been developed to study biological membranes without interferences from cell machinery. To obtain a smooth hydrophilic biomembrane
support and a mode of binding of the membrane, either a crosslinked gel or an aromatic polyamine-polymer doped with avidin
was deposited at the metal electrode by electropolymerization. This layer (less than 10 nm thick) also served as a submembrane
compartment. The facilitated glucose transporter (GLUT-1) purified from human erythrocytes was integrated into a lipid membrane
containing artificial biotinylated lipids and reacted with the activated surface of the glucose sensitive electrode. It was
demonstrated that the lipid layer was attached to the polymer-containing avidin and could only be removed by detergent extraction.
The presence of an active membrane transporter was demonstrated by electrochemical detection of glucose in the submembrane
compartment, and by inhibition of glucose transport with the specific inhibitor Cytochalasin-B. 相似文献
64.
A cobaltocenium label was covalently attached to two antidepressants, nortriptyline and desipramine, via an amide linkage, and also to the hydrazine derivative of the biologically important compound biotin (vitamin H), again via an amide linkage. Analytically pure samples of these new cobaltocenium salts could be obtained by chromatography on silica gel followed by elution with aqueous acetone solutions containing sodium chloride (NaCl). These positively charged cobaltocenium ions preconcentrate in a polyanionic Nafion film coated on a glassy carbon surface, albeit at different concentration levels. One factor which seems to influence the amount of cobaltocenium ion that enters the film is polarity since the cobaltocenium ion containing the rather polar biotin preconcentrates at the lowest level in the relatively hydrophobic Nafion. Square-wave voltammograms of Nafion films containing these cobaltocenium cations exhibit a one-electron, reversible, reduction wave at approximately ?1.1 V (vs Ag/AgCl) with peak currents that are sufficiently large to permit detection of 10?8 M quantities of these substances in the bulk solution. 相似文献
65.
《Analytical letters》2012,45(10):2045-2065
Abstract The research work reported herein is the development of a simple and specific quantitative procedure for the determination of P. falciparum DNA in malaria that involves the direct detection of the highly 42‐kDa conserved C‐terminal regiopn of P. falciparum merozoite surface protein gene (MSP1 42 gene). This procedure entails the amplification of the MSP1 42 gene by using the PCR technique in the presence of digoxigenin‐11‐dUTP and the synthesis of the specific biotin label nucleotide probes directed to the MSP1 42 gene. These specific probes are then used in the Enzyme Linked Immunosorbent Assay (ELISA) for the quantitative determination of the MSP1 42 gene which leads to the quantitative determination of P. falciparum DNA in malaria for quantitative diagnostic purpose. The P. falciparum malaria diagnostic results obtained from a small number of 18 whole blood samples show that the present quantitative PCR‐ELISA procedure allows the quantitative determination of P. falciparum DNA in malaria with a sensitivity and specificity over to those of the current standard microscopic examination. This quantitative PCR‐ELISA procedure is not only important for quantitative P. falciparum malaria diagnosis but also useful for monitoring the efficacy of any existing anti‐malarial drug as well as for testing the efficacy of any malaria vaccine. 相似文献
66.
67.
Electropolymerization as a versatile route for immobilizing biological species onto surfaces 总被引:5,自引:0,他引:5
Bidan G Billon M Galasso K Livache T Mathis G Roget A Torres-Rodriguez LM Vieil E 《Applied biochemistry and biotechnology》2000,89(2-3):183-193
Biosensors based on electronic conducting polymers appear particularly well suited to the requirements of modern biological
analysis—multiparametric assays, high information density, and miniaturization. We describe a new methodology for the preparation
of addressed DNA matrices. The process includes an electrochemically directed copolymerization of pyrrole and oligonucleotides
bearing on their 5′ end a pyrrole moiety. The resulting polymer film deposited on the addressed electrode consists of pyrrole
chains bearing covalently linked oligonucleotides (ODN). An oligonucleotide array was constructed on a silicon device bearing
a matrix of 48 addressable 50 × 50 μm gold microelectrodes. This technology was successfully applied to the genotyping of
hepatitis C virus in blood samples. Fluorescence detection results show good sensitivity and a high degree of spatial resolution.
In addition, gravimetric studies carried out by the quartz crystal microbalance technique provide quantitative data on the
amount of surface-immobilized species. In the case of ODN, it allows discrimination between hybridization and nonspecific
adsorption. The need for versatile processes for the immobilization of biological species on surfaces led us to extend our
methodology. A biotinylated surface was obtained by coelectropolymerization of pyrrole and biotin-pyrrole monomers. The efficiency
for recognition (and consequently immobilization) of R-phycoerythrin-avidin was demonstrated by fluorescence detection. Copolymerization
of decreasing ratios of pyrrole-biotin over pyrrole allowed us to obtain a decreasing scale of fluorescence. 相似文献
68.
69.
XU Yechun SHEN Jianhua LUO Xiaomin SHEN Xu CHEN Kaixian & JIANG Hualiang Center for Drug Discovery Design State Key Laboratory of New Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences ChineseAcademy of Sciences Shanghai China 《中国科学B辑(英文版)》2004,47(5):355-366
Molecular recognition and specific protein-ligandinteractions are central to many biochemical processes,such as enzyme catalysis, assembly of organelles, en-ergy transduction, signaling, diverse control functions,and replication, expression and storage of the geneticmaterial[1]. Moreover, protein-ligand interactions pro-vide the mechanism of many drug therapies and un-derstanding of such interactions is thus significant forrational drug design[1,2]. For the experimental studiesof protein-ligan… 相似文献
70.
Cosnier S 《Applied biochemistry and biotechnology》2000,89(2-3):127-138
The concept and potentialities of electrochemical procedures of biomolecule immobilization are described. The entrapment of
biomolecules within electropolymerized films consists of the application of an appropriate potential to an electrode soaked
in an aqueous solution containing monomer and biomolecules. This method of biosensor construction is compared with a two-step
procedure based on the adsorption of an aqueous amphiphilic pyrrole monomer-biomolecule mixture on an electrode followed by
the electropolymerization of the adsorbed monomers. Another approach is based on the electrogeneration of polymer films functionalized
by specific groups allowing subsequently the attachment of biomolecules. The immobilization of biomolecules on these films
by covalent binding or noncovalent interactions is described. 相似文献