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61.
Drees BE Weipert A Hudson H Ferguson CG Chakravarty L Prestwich GD 《Combinatorial chemistry & high throughput screening》2003,6(4):321-330
We describe the development and implementation of competitive fluorescence polarization (FP) based assays for determining activity of phosphoinositide 3-kinase (PI 3-K) and the type-II SH2-domain-containing inositol 5-phosphatase (SHIP2). These assays are based on the interaction of specific phosphoinositide binding proteins with fluorophore-labeled phosphoinositide and inositol phosphate tracers. Enzyme reaction products are detected by their ability to compete with the fluorescent tracers for protein binding, leading to an increase in the amount of free tracer and a decrease in polarization (mP) values. A variety of fluorophore-labeled tracers were evaluated, and assay sensitivity and specificity for products of PI 3-K and SHIP2 activity was determined. Assay performance was evaluated using recombinant PI 3-Kalpha and SHIP2 with diC(8)-PI(4,5)P(2) and diC(8)-PI(3,4,5)P(3) as respective substrates. IC(50) values for previously characterized PI 3-K inhibitors were within expected ranges. These assays are homogeneous, sensitive, and rapid, and suitable for HTS applications, and will facilitate screening for novel inhibitors of phosphoinositide kinases and phosphatases in drug development. 相似文献
62.
We report here a convenient and efficient two-step synthesis of 1-aryl-2-arylamino-4-alkyl/phenyl-5-aroyl-1H-imidazoles from easily available amidinothioureas. Guanylation of amidinothioureas 1 using mercury(II) chloride as a thiophile yielded amidinoguanidines 2, which reacted with various phenacyl bromides under mild conditions to afford the corresponding diversely functionalized imidazoles 3 in moderate to good yields. 相似文献
63.
Many well-established homogeneous separation free immunoassays rely on particulate label technologies. Particles generally contain a high concentration of the embedded label and they have a large surface area, which enables conjugation of a large amount of protein per particle. Eu(III)-chelate dyed nanoparticles have been successfully used as labels in heterogeneous and homogeneous immunoassays. In this study, we compared the characteristics of two homogeneous competitive immunoassays using either soluble Eu(III)-chelates or polystyrene particles containing Eu(III)-chelates as donors in a fluorescence resonance energy transfer based assay. The use of the particulate label significantly increased the obtained sensitized emission, which was generated by a single binding event. This was due to the extremely high specific activity of the nanoparticle label and also in some extent the longer Förster radius between the donor and the acceptor. The amount of the binder protein used in the assay could be decreased by 10-fold without impairing the obtainable sensitized emission, which subsequently led to improved assay sensitivity. The optimized assay using particulate donor had the lowest limit of detection (calculated using 3 × S.D. of the 0 nM standard) 50 pM of estradiol in the assay well, which was approximately 20-fold more sensitive than assays using soluble Eu(III)-chelates. 相似文献
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Revisiting thiol-yne chemistry: Selective and efficient monoaddition for block and graft copolymer formation
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Johannes K. Sprafke Jason M. Spruell Kaila M. Mattson Damien Montarnal Alaina J. McGrath Robert Pötzsch Daigo Miyajima Jerry Hu Allegra A. Latimer Brigitte I. Voit Takuzo Aida Craig J. Hawker 《Journal of polymer science. Part A, Polymer chemistry》2015,53(2):319-326
The untapped potential of radical thiol-yne mono-addition chemistry is exploited to overcome the known limitations of thiol-ene chemistry in polymer coupling and block copolymer formation. By careful choice of alkyne, the reaction can selectively lead to the mono-addition product with efficiencies surpassing those achieved by traditional thiol-ene chemistry. This improvement is illustrated by the nearly quantitative synthesis of a variety of diblock and graft copolymers. © 2014 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2015 , 53, 319–326 相似文献
67.
Fei Fei Soo-Mi Kweon Leena Haataja Paulo De Sepulveda John Groffen Nora Heisterkamp 《BMC biochemistry》2010,11(1):48
Background
RhoGDI proteins are important regulators of the small GTPase Rac, because they shuttle Rac from the cytoplasm to membranes and also protect Rac from activation, deactivation and degradation. How the binding and release of Rac from RhoGDI is regulated is not precisely understood. 相似文献68.
Jitendra C. Kaila 《Tetrahedron letters》2010,51(11):1486-1489
A thiophile-promoted one-pot synthesis of trisubstituted 1,3,5-triazines starting from isothiocyanates, N,N-diethylamidines, and carbamidines has been studied. The reaction proceeds through the formation of intermediary amidinothioureas, which react with carbamidines in the presence of mercury(II) chloride to generate the desired 1,3,5-triazines in good to moderate yields (40-70%). 相似文献
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Szajnman SH Rosso VS Malayil L Smith A Moreno SN Docampo R Rodriguez JB 《Organic & biomolecular chemistry》2012,10(7):1424-1433
α-Fluorinated-1,1-bisphosphonic acids derived from fatty acids were designed, synthesized and biologically evaluated against Trypanosoma cruzi, the etiologic agent of Chagas disease, and against Toxoplasma gondii, the agent responsible for toxoplasmosis, and also towards the target parasitic enzymes farnesyl pyrophosphate synthase of T. cruzi (TcFPPS) and T. gondii (TgFPPS). Interestingly, 1-fluorononylidene-1,1-bisphosphonic acid (compound 43) proved to be an extremely potent inhibitor of the enzymatic activity of TgFPPS at the low nanomolar range, exhibiting an IC(50) of 30 nM. This compound was two-fold more potent than risedronate (IC(50) = 74 nM) that was taken as a positive control. This enzymatic activity was associated with a strong cell growth inhibition against tachyzoites of T. gondii, with an IC(50) value of 2.7 μM. 相似文献