全文获取类型
收费全文 | 1095篇 |
免费 | 72篇 |
国内免费 | 21篇 |
出版年
2023年 | 8篇 |
2022年 | 19篇 |
2021年 | 24篇 |
2020年 | 24篇 |
2019年 | 31篇 |
2018年 | 28篇 |
2017年 | 19篇 |
2016年 | 36篇 |
2015年 | 53篇 |
2014年 | 48篇 |
2013年 | 60篇 |
2012年 | 40篇 |
2011年 | 65篇 |
2010年 | 38篇 |
2009年 | 49篇 |
2008年 | 51篇 |
2007年 | 51篇 |
2006年 | 49篇 |
2005年 | 45篇 |
2004年 | 47篇 |
2003年 | 34篇 |
2002年 | 32篇 |
2001年 | 17篇 |
2000年 | 20篇 |
1999年 | 21篇 |
1998年 | 17篇 |
1997年 | 12篇 |
1996年 | 8篇 |
1995年 | 8篇 |
1994年 | 6篇 |
1993年 | 18篇 |
1992年 | 11篇 |
1991年 | 7篇 |
1990年 | 9篇 |
1989年 | 14篇 |
1988年 | 12篇 |
1987年 | 10篇 |
1986年 | 9篇 |
1985年 | 9篇 |
1984年 | 29篇 |
1983年 | 22篇 |
1982年 | 20篇 |
1981年 | 18篇 |
1980年 | 11篇 |
1979年 | 6篇 |
1978年 | 4篇 |
1977年 | 5篇 |
1975年 | 3篇 |
1974年 | 3篇 |
1973年 | 4篇 |
排序方式: 共有1188条查询结果,搜索用时 15 毫秒
71.
Macroaffinity ligand-facilitated three-phase partitioning (MLFTPP) for purification of xylanase 总被引:2,自引:0,他引:2
It is shown that eudragit S-100, a copolymer of methylacrylic acid and methylmethacrylate, undergoes three-phase partitioning. It was found that 95% eudragit S-100 could be recovered as the interfacial precipitate by using 30% (w/v) ammonium sulfate, 1:1 ratio of t-butanol to polymer solution at 40 degrees C. Three-phase partitioning of proteins uses simultaneous addition of ammonium sulfate and t-butanol to precipitate proteins in an interfacial layer separating the aqueous phase and organic solvent. Exploiting the affinity of xylanases towards eudragit S-100, it was possible to purify xylanase from Aspergillus niger; 60% recovery of activity with 95-fold purification could be obtained by this process. The purified enzyme showed A single band on SDS-PAGE. The technique shows promise to develop into a general method that could be termed "macroaffinity ligand-facilitated three-phase partitioning (MLFTPP). 相似文献
72.
73.
74.
75.
Sarnataro D Paladino S Campana V Grassi J Nitsch L Zurzolo C 《Traffic (Copenhagen, Denmark)》2002,3(11):810-821
PrP(C) is a glycosylphosphatidylinositol-anchored protein expressed in neurons as well as in the cells of several peripheral tissues. Although the normal function of PrP(C) remains unknown, a conformational isoform called PrP(Sc) (scrapie) has been proposed to be the infectious agent of transmissible spongiform encephalopathies in animals and humans. Where and how the PrP(C) to PrP(Sc) conversion occurs in the cells is not yet known. Therefore, dissecting the intracellular trafficking of the wild-type prion protein, as well as of the scrapie isoform, can be of major relevance to the pathogenesis of the diseases. In this report we have analyzed the exocytic pathway of transfected mouse PrP(C) in thyroid and kidney polarized epithelial cells. In contrast to the majority of glycosylphosphatidylinositol-anchored proteins, we found that PrP(C) is localized mainly on the basolateral domain of the plasma membrane of both cell lines. This is reminiscent of the predominant somatodendritic localization found in neurons. However, similarly to apical glycosylphosphatidylinositol-proteins, PrP(C) associates with detergent-resistant microdomains, which have been suggested to have a role in apical sorting of glycosylphosphatidylinositol-proteins, as well as in the conversion process of PrP(C) to PrP(Sc). In order to discriminate whether detergent-resistant microdomains have a direct role in PrP(Sc) conversion, or whether they are involved in the transport of the protein to the site of its conversion, we have examined the effect of disruption of detergent-resistant microdomain association on PrP(C) intracellular traffic. Consistent with the unusual basolateral localization of this glycosylphosphatidylinositol-linked protein, our data exclude a classical role for detergent-resistant microdomains in the post-trans-Golgi network sorting and transport of PrP(C) to the plasma membrane. 相似文献
76.
Rambotti MG Spreca A Giambanco I Sorci G Donato R 《Molecular and cellular biochemistry》2002,230(1-2):85-96
Membrane-bound guanylate cyclase activity was detected by ultracytochemistry at the electron microscope level in several mammalian tissues. The technique used in these studies allows the detection of active enzyme at the membrane site where it is located. In a few cases, such as normal and regenerating peripheral nerves and placenta, membrane-bound guanylate cyclase could be detected in the absence of stimulators of enzyme activity. However, in the majority of these studies membrane-bound guanylate cyclase was investigated following stimulation with natriuretic peptides, guanylin, or the Ca2+ sensor proteins, S100B and S100A1. In general, membrane-bound guanylate cyclase was localized to plasma membranes, in accordance with the functional role of this enzyme. Yet, in secretory cells the enzyme activity was localized on intracellular membranes, suggesting a role of membrane-bound guanylate cyclase in secretory processes. Finally, S100B and S100A1 were found to colocalize with membrane-bound guanylate cyclase on photoreceptor disc membranes and to stimulate enzyme activity at these sites in dark-adapted retinas in a Ca2+-dependent manner. The results of these analyses are discussed in relation to the proposed functional role(s) of this enzyme. 相似文献
77.
Sekher Pannala A Bruckdorfer KR Rice-Evans CA 《Biochemical and biophysical research communications》2002,292(1):175-183
Previous studies have demonstrated increased plasma levels of oxidised low-density lipoprotein (oxLDL) in chronic smokers, which has been associated with the extent of endothelial dysfunction. In this study we examine the relationship between the amino acid composition of apolipoprotein B100 (apo B) of low-density lipoprotein (LDL), by reverse phase HPLC after precolumn derivatisation, between smokers (> or =40 cigarettes/day) and nonsmokers in relation to their plasma and LDL antioxidant status. While there was a significant difference in the levels of plasma vitamin C and alpha-tocopherol between female smokers and nonsmokers, as well as in the levels of LDL alpha-tocopherol, there was no significant difference in the amino acid composition of apo B between the two groups. 相似文献
78.
Kizawa K Troxler H Kleinert P Inoue T Toyoda M Morohashi M Heizmann CW 《Biochemical and biophysical research communications》2002,299(5):857-862
S100A3, a unique protein among all members of the calcium-binding S100 family, is specifically expressed at the inner endocuticle of human hair fibers. Upon hair damage, S100A3 is released from hair fibers and possibly destabilizes the hair tissue architecture. This study describes the purification and characterization of native S100A3 isolated from human hair fibers. We extracted native S100A3 from cuticles and purified the protein by anion-exchange chromatography. The results of 2D gel electrophoresis showed that cuticle S100A3 has a slightly lower isoelectric point compared to the recombinant protein. Tandem mass spectrometry of the peptides resulting from endoproteinase digest of cuticle S100A3 revealed that the N-terminal methionine is replaced with an acetyl group. This is the first report on biochemical characteristics of S100A3 in hair cuticle. 相似文献
79.
Tyrosine phosphorylation of a novel 100-kDa protein coupled to CD28 in resting human T cells is enhanced by a signal through TCR/CD3 complex 总被引:1,自引:0,他引:1
Matsumoto A Dobashi H Ohnishi H Tanaka T Kubota Y Kitanaka A Ishida H Tokuda M Waki M Kubo A Ishida T 《Microbiology and immunology》2003,47(1):63-69
For T cell activation, two signals are required, i.e., a T cell receptor (TCR)/CD3-mediated main signal and a CD28-mediated costimulatory signal. CD28 binds to its ligand (CD80 or CD86) and transduces the most important costimulatory signal. The cytoplasmic domain of the CD28 molecule, composed of 41 amino acids, does not contain any intrinsic enzyme activity. The cytoplasmic domain of CD28 is remarkably conserved among species and is associated with a number of signaling molecules that affect the main signal. We report here that a tyrosine phosphorylated 100-kDa protein (ppl00) was coupled to the CD28 cytoplasmic domain in Jurkat and human peripheral T cells. The pp100 was distinguished from other CD28 associated molecules such as Vav, STAT5, PI 3-kinase, Valosin-containing protein (VCP), Nucleolin, Gab2 (Grb2-associated binding protein 2), and STAT6. The tyrosine phosphorylation of pp100 coprecipitated with CD28 was enhanced by CD3 stimulation by the specific antibody, tyrosine phosphatase inhibitor and PKC activator. Tyrosine phosphorylation of pp100 was attenuated by the prior addition of PKC inhibitor. These findings indicate that pp100 is a novel tyrosine phosphorylated protein coupled to CD28 under continuous control of tyrosine phosphatases and might play a role in T cell activation augmented by a TCR/CD3-mediated main signal. 相似文献
80.
Recovery of the proteose peptone component 3 from cheese whey was optimal using a 16% (w/w) Reppal PES 100 – 24% (w/w) PEG 600 aqueous two-phase system, at pH 7, giving a mass recovery yield of 99% and a purity of 83% for proteose peptone component 3 in the upper phase. Using the above system a partition coefficient of 30.7 and a purification factor of 6.9 were achieved. 相似文献