全文获取类型
收费全文 | 453篇 |
免费 | 12篇 |
国内免费 | 20篇 |
出版年
2023年 | 3篇 |
2022年 | 4篇 |
2021年 | 7篇 |
2020年 | 4篇 |
2019年 | 12篇 |
2018年 | 5篇 |
2017年 | 7篇 |
2016年 | 3篇 |
2015年 | 12篇 |
2014年 | 27篇 |
2013年 | 25篇 |
2012年 | 16篇 |
2011年 | 24篇 |
2010年 | 27篇 |
2009年 | 31篇 |
2008年 | 35篇 |
2007年 | 26篇 |
2006年 | 24篇 |
2005年 | 26篇 |
2004年 | 27篇 |
2003年 | 20篇 |
2002年 | 15篇 |
2001年 | 9篇 |
2000年 | 9篇 |
1999年 | 8篇 |
1998年 | 11篇 |
1997年 | 6篇 |
1996年 | 4篇 |
1995年 | 6篇 |
1994年 | 8篇 |
1993年 | 6篇 |
1992年 | 4篇 |
1991年 | 2篇 |
1990年 | 4篇 |
1989年 | 4篇 |
1988年 | 1篇 |
1987年 | 4篇 |
1986年 | 2篇 |
1985年 | 4篇 |
1983年 | 3篇 |
1982年 | 3篇 |
1981年 | 3篇 |
1980年 | 1篇 |
1977年 | 1篇 |
1976年 | 2篇 |
排序方式: 共有485条查询结果,搜索用时 563 毫秒
1.
Trigonopsis variabilis induced for D-amino acid oxidase and catalase was immobilized by entrapment in Polyacrylamide beads obtained by radiation
polymerisation. Permeabilization of the cells was found to be essential for optimal activity of the enzymes in free cells.
However, the process of entrapment itself was found to eliminate the permeability barrier of cells immobilized in Polyacrylamide.
The two enzymes exhibited a differential response on Polyacrylamide entrapment. Thus, D-amino acid oxidase activity was stabilized
to heat inactivation whereas catalase in the same cells showed a destabilization on entrapment in Polyacrylamide. The coimmobilized
enzyme preparation showed an operational half life of 7–9 days after which the D-amino acid oxidase activity remained stable
at a value 35–40% of that of the initial activity for a study period of 3 weeks. Coimmobilization of MnO2 was not effective in enhancing the operational life of the enzyme preparation. 相似文献
2.
Size selection of latex beads by blackfly larvae (Diptera: Simuliidae) in the laboratory 总被引:1,自引:1,他引:0
P. Schröder 《Hydrobiologia》1987,144(2):163-171
In laboratory experiments, blackfly larvae collected from a lake outlet, a woodland and a meadow stream were tested for size
selection of latex beads of < Ito > 100 μm diameters. 3 suspensions of varying proportions for each size category were supplied
to these blackfly larvae in common experiments. Comparisons between the size frequency distributions of particles supplied
and the particle compositions in the larval guts showed intra- and interspecific differences and were quantified by calculating
Jacobs' electivity index. In all species selection of larger particles increased with the larger larval instars. Although
there was a positive selectivity of small particles in some cases, the ingested proportion of large particles increases volumes
and biomasses of gut content and may be more important for larval growth than small particles. 相似文献
3.
Françoise Farace Anne-Marie Le Ridant Bernard Escudier Thierry Hercend Frédéric Triebel 《Biotherapy》1992,5(1):11-20
We have attempted to improve negative selection procedures for the large scale purification of human CD
in3
–
CD56+ NK cells. In a series of experiments, purifications of NK cells from 108 PBMC were performed by T cell depletion using either direct or indirect anti-CD3 labeling and the Magnetic Activated Cell Separation (MACS) procedure. Contaminating CD3+ cells were still present using either one of these two different T cell depletion protocols as shown by phenotyping IL-2 supplemented cell cultures on day 12. A second cycle of purification was therefore added. When MACS and Dynabeads were compared as complementary procedures to the first MACS cycle starting with 108 cells, the Dynabeads method was found to be superior to the MACS with regard to the elimination of residual T cells. Starting from 109 PBMC, we showed that this MACS+Dynabeads procedure gave similar satisfactory results when compared to the scaling-up of a previously established two steps procedure using Dynabeads. These two approaches (MACS+Dynabeads and 2 cycles of Dynabeads) have been also tested in a clinical setting to purify NK cells from cancer patients prior toin vitro expansion. The results indicate that the two methods are equivalent with respect to purity and recovery rate; a slight advantage in terms of feasibility was found in favor of 2 cycles of Dynabeads. 相似文献
4.
Mortimer M. Civan Stephen R. Williams David G. Gadian Enrique Rozengurt 《The Journal of membrane biology》1986,94(1):55-64
Summary Swiss mouse 3T3 cells grown on microcarrier beads were superfused with electrolyte solution during continuous NMR analysis. Conventional31P and19F probes of intracellular pH (pH
c
) were found to be impracticable. Cells were therefore superfused with 1 to 4mm 2-deoxyglucose, producing a large intracellular, pH-sensitive signal of 2-deoxyglucose phosphate (2DGP). The intracellular incorporation of 2DGP inhibited the Embden-Meyerhof pathway. However, intracellular ATP was at least in part retained and the cellular responsivity to changes in extracellular ionic composition and to the application of growth factors proved intact. Transient replacement of external Na+ with choline or K+ reversibly acidified the intracellular fluids. Quiescent cells and mitogenically stimulated cells displayed the same dependence of shifts in pH
c
on external Na+ concentration (c
Na
o
). pH
c
also depended on intracellular Na+ concentration (c
Na
o
). Increasingc
Na
c
by withdrawing external K+ (thereby inhibiting the Na,K-pump) caused reversible intracellular acidification; subsequently reducingc
Na
o
produced a larger acid shift in pH
c
than with external K+ present. Comparison of separate preparations indicated that pH
c
was higher in stimulated than in quiescent cells. Transient administration of mitogens also reversibly alkalinized quiescent cells studied continuously. This study documents the feasibility of monitoring pH
c
of Swiss mouse 3T3 cells using31P NMR analysis of 2DGP. The results support the concept of a Na/H antiport operative in these cells, both in quiescence and after mitogenic stimulation. The data document by an independent technique that cytoplasmic alkalinization is an early event in mitogenesis, and that full activity of the Embden-Meyerhof pathway is not required for the expression of this event. 相似文献
5.
经N-乙酰氨基葡萄糖交联琼脂糖亲和层析及以交联琼脂糖介质的高效液相分子筛层析,从中国鲎细胞溶解物中分离纯化了一种凝集素,其活性比原料鲎试剂提高128倍。鲎凝集素SDS电泳时表现出分子量为69000,和72000的二个亚基。N-乙酰氨基葡萄糖、D-半乳糖,D-甘露糖及岩藻糖等对鲎凝集素凝集鸡红细胞的活性有显著抑制作用,加热60℃,10分钟可使凝集素活性基本丧失。CaCl_2为凝集素活性所必需。鲎凝集素与肺炎球菌C多糖有沉淀反应。 相似文献
6.
Kristel Eggermont Inge J. Goderis Willem F. Broekaert 《Plant Molecular Biology Reporter》1996,14(3):273-279
We describe here a reliable high-throughput method for extraction of RNA from fresh or frozen plant tissue that obviates laborious
and time-consuming homogenisation by mortar and pestle. The method is based on homogenisation by high-speed reciprocal shaking
in presence of a mixture of inexpensive abrasive materials; i.e., quartz sand and glass beads. After homogenisation, the method
follows a standard procedure for RNA extraction by phenol/LiCl. Yield and quality of RNA obtained by homogenisation with the
sand/glass bead mix are identical to those obtained by mortar and pestle. 相似文献
7.
The efficiency of lgM production by hybridoma cells (1) cultured in suspension; (2) entrapped in alginate beads; or (3) packed in hollowfiber cartridge bioreactors, were compared in long-term perfusion cultures. The results showed that steady-state cell concentration and antibody production, per liter of perfused medium per day, were similar when cells were either entrapped in alginate beads of maintained in suspension. These values were also similar whether cells were maintained at high density in a hollowfiber cartridge bioreactro, or at low density in suspension. This work points out that cell behavior and antibody yield are comparable overall in the various perfusion systems currently used. However, a significant reduction of antibody production appeared whenever a part of the viable cells was lost in the filtrate. The reduction was due both to a decrease of viable cell yield and a decline of lgM productivity on a percell basis. This result is well in agreement with the previously presented model of "grow or die" cell cycle system of hybridoma, which proposes that the ratio of arrested to proliferating cells in perfusion cultures, should be increased in proportion to cell retention in the bioreactor, with a concomitant increase of lgM productivity. 相似文献
8.
Cellulose-based stationary phases are known to be very efficient and versatile chiral sorbents for the chromatographic resolution of racemates. Except for microcrystalline cellulose triacetate (CTA I), basically all other cellulose-based phases have been prepared by coating of ca. 20% weight polymer on a wide pore silica gel used as a carrier. In this work we describe the preparation of benzoylcellulose (TBC) beads in the pure polymeric form (without inorganic carrier) from an emulsion of the organic polymer. The new material has been fully characterized and used as a chiral stationary phase for the resolution of various classes of racemic compounds such as benzylic alcohols or acetate derivatives of aliphatic alcohols and diols. The structural variety of the separated solutes as well as the irrational influence of the aromatic substituent in different classes of aryl compounds suggest that multiple interaction sites are involved in the complexation, making a prediction of the separation difficult. The benzoyl cellulose beads exhibit a very high loading capacity, which is particularly useful for preparative purposes as demonstrated for selected examples. 相似文献
9.
The suitability of bead mills for the release of intracellular bacterial enzymes has been studied using the Dyno-Mill Model KDL. The effect of cell concentration, bead size and agitation speed on the release of beta-lactamase from Enterobacter cloacae P99 was examined. Scale-up considerations included, the best operational values for these parameters were 1 g cell paste suspended in 2.5 ml buffer, 0.25 mm diameter glass beads and 15 ms −1 agitation speed. These conditions proved suitable for the release of enzymes from other Gram-negative bacteria in both batch and continuous processes. 相似文献
10.
Adegoke Isiaka Adetunji 《Preparative biochemistry & biotechnology》2013,43(10):898-905
AbstractExtracellular lipase from an indigenous Bacillus aryabhattai SE3-PB was immobilized in alginate beads by entrapment method. After optimization of immobilization conditions, maximum immobilization efficiencies of 77%?±?1.53% and 75.99%?±?3.49% were recorded at optimum concentrations of 2% (w/v) sodium alginate and 0.2?M calcium chloride, respectively, for the entrapped enzyme. Biochemical properties of both free and immobilized lipase revealed no change in the optimum temperature and pH of both enzyme preparations, with maximum activity attained at 60?°C and 9.5, respectively. In comparison to free lipase, the immobilized enzyme exhibited improved stability over the studied pH range (8.5–9.5) and temperature (55–65?°C) when incubated for 3?h. Furthermore, the immobilized lipase showed enhanced enzyme-substrate affinity and higher catalytic efficiency when compared to soluble enzyme. The entrapped enzyme was also found to be more stable, retaining 61.51% and 49.44% of its original activity after being stored for 30 days at 4?°C and 25?°C, respectively. In addition, the insolubilized enzyme exhibited good reusability with 18.46% relative activity after being repeatedly used for six times. These findings suggest the efficient and sustainable use of the developed immobilized lipase for various biotechnological applications. 相似文献