全文获取类型
收费全文 | 1224篇 |
免费 | 25篇 |
国内免费 | 52篇 |
出版年
2023年 | 6篇 |
2022年 | 9篇 |
2021年 | 18篇 |
2020年 | 14篇 |
2019年 | 20篇 |
2018年 | 22篇 |
2017年 | 37篇 |
2016年 | 55篇 |
2015年 | 55篇 |
2014年 | 73篇 |
2013年 | 72篇 |
2012年 | 53篇 |
2011年 | 65篇 |
2010年 | 67篇 |
2009年 | 51篇 |
2008年 | 51篇 |
2007年 | 77篇 |
2006年 | 53篇 |
2005年 | 45篇 |
2004年 | 41篇 |
2003年 | 42篇 |
2002年 | 25篇 |
2001年 | 11篇 |
2000年 | 19篇 |
1999年 | 20篇 |
1998年 | 16篇 |
1997年 | 20篇 |
1996年 | 21篇 |
1995年 | 26篇 |
1994年 | 38篇 |
1993年 | 17篇 |
1992年 | 11篇 |
1991年 | 16篇 |
1990年 | 6篇 |
1989年 | 23篇 |
1988年 | 9篇 |
1987年 | 11篇 |
1985年 | 5篇 |
1984年 | 8篇 |
1983年 | 11篇 |
1982年 | 12篇 |
1981年 | 12篇 |
1980年 | 12篇 |
1979年 | 6篇 |
1978年 | 1篇 |
1977年 | 1篇 |
1976年 | 4篇 |
1975年 | 4篇 |
1974年 | 3篇 |
1973年 | 6篇 |
排序方式: 共有1301条查询结果,搜索用时 15 毫秒
1.
Antisera were raised against L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine (DA), noradrenaline (NA), and octopamine (OA). This was achieved by coupling each molecule to bovine serum albumin or human serum albumin using glutaraldehyde. The conjugated aromatic amines were kept in a reducing medium containing sodium metabisulfite. Antiserum specificity was tested using an enzyme-linked immunosorbent assay method for catecholamines. Competition experiments were done between the immunogen coated on the well plates and each catecholamine, either in the free state or in conjugated form, previously incubated with an antiserum. In each case, the nonconjugated compound was poorly recognized. The nonreduced conjugates of L-DOPA and DA were well recognized, whereas those of NA and OA were poorly immunoreactive. The cross-reactivity ratios established in the competition experiments allowed the specificity of the immune response to be defined. In each case, it was found to be high. The results suggest that the antibodies of L-DOPA and DA antisera recognize preferentially the catechol moiety, whereas for the anti-NA and anti-OA antibodies, the lateral chain is important. 相似文献
2.
Summary Mineral transport across the plasma membrane of plant cells is controlled by an electrochemical gradient of protons. This
gradient is generated by an ATP-consuming enzyme in the membrane known as a proton pump, or H+-ATPase. The protein has a catalytic subunit of Mr=100,000 and is a prominent band when plasma membrane proteins are analyzed
by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
We generated specific rabbit polyclonal antibody against the Mr=100,000 H+-ATPase and used the antibody to screen λgtll expression vector libraries of plant DNA. Several phage clones producing immunoreactive
protein, and presumably containing DNA sequences for the ATPase structural gene, were isolated and purified from a carrot
cDNA library and a Arabidopsis genomic DNA library. These studies represent our first efforts at cloning the structural gene
for a plant plasma membrane transport protein. Applicability of the technique to other transport protein genes and the potential
for use of recombinant DNA technology in plant mineral transport research are discussed. 相似文献
3.
4.
Polyclonal antibodies were prepared against the purified elongation factor Tu (EF-Tu) of Escherichia coli and Bacillus subtilis. Using the methods of Western blotting and microcomplement fixation the cross-reactivities of EF-Tu of 19 different prokaryotes were determined. The immunological distance were compared with the results of 16S rRNA oligonucleotide analysis. An unexpectedly high cross-reactivity was revealed between the EF-Tu of B. subtilis and the antiserum against the EF-Tu of E. coli. A comparison of the predicted amino acid sequences from the tuf-genes of E. coli and B. subtilis yielded two identical peptide fragments that are likely candidates for antibody binding sites.Abbreviations EF-Tu
elongation factor Tu
- GDP
guanosine 5-diphosphate
- GTP
guanosine 5-triphosphate
- MCF
microcomplement fixation
- T
type strain 相似文献
5.
Richard M. Napier Michael A. Venis Michael A. Bolton Louise I. Richardson Goeffrey W. Butcher 《Planta》1988,176(4):519-526
Binding proteins, thought to be auxin receptors, can be solubilised from maize (Zea mays L.) membranes after acetone treatment. From these crude extracts, receptor preparations of over 50% purity can be obtained by a reliable, straight-forward procedure involving three chromatographic steps — anion exchange, gel filtration and high-resolution anion exchange. Such preparations have been used to immunise rats for subsequent production of monoclonal antibodies. By the further step of native polyacrylamide gel electrophoresis the semi-purified preparations yield homogeneous, dimeric (22-kilodalton, kDa) auxin-binding protein, which has been used to produce a polyclonal rabbit antiserum. The preliminary characterisation of this antiserum and of the five monoclonal antibodies is presented. Two of the monoclonal antibodies specifically recognise the major 22-kDa-binding protein polypeptide whilst the other three recognise, in addition, a minor 21-kDa species. All the monoclonal antibodies recognise the polypeptide rather than the glycan side chain and the polyclonal antiserum also recognises deglycosylated binding protein. The antibodies have been used to quantify the abundance of auxinbinding protein in a number of tissues of etiolated maize seedlings. Root membranes contain 20-fold less binding protein than coleoptile membranes.Abbreviations ABP
auxin-binding protein
- DEAE
diethylaminoethyl
- Ig
immunoglobulin
- kDa
kilodalton
- NAA
naphthalene-1-acetic acid
- Mr
relative molecular mass
- PAGE
polyacrylamide gel electrophoresis
- SDS
sodium dodecyl sulfate 相似文献
6.
John Greenman Nancy Hogg Suzanne Nikoletti Christopher Slade George Stevenson Martin Glennie 《Cancer immunology, immunotherapy : CII》1992,34(6):361-369
Summary The three forms of Fc receptor carried by monocytes (FcRI, II) and natural killer (NK) cells (FcRIII) are all capable of mediating cell lysis. Here we compare the use of F(ab)2 bispecific antibodies, specifically targetting individual FcR, and chimeric IgG mouse/human antibodies which are capable of targetting all FcR, for their ability to mediate target cell destruction. The derivatives are prepared by linking hinge sulphydryl residues via tandem thioether bonds, using a bismaleimide crosslinker: Fab from an anti-FcR mAb linked to Fab from a common anti-target mAb (BsAb), or Fab from the common anti-target mouse antibody linked to human Fc (FabFc or bisFabFc). All the derivatives targetting chick red blood cells gave efficient lysis, although different effector cell donors yielded differences in both the lytic levels achieved and the comparative efficiencies of derivatives. In contrast, significant lysis of the guinea pig lymphoblastic leukaemia, L2C, regularly resulted only via the anti-FcRIII BsAb and the chimeric derivatives. These results suggest that the chimeric, Fc-containing derivatives mediate tumour cell lysis principally through FcRIII on NK cells. This is in contrast to the situation with the chick red blood cells where the chimeric derivatives appear capable of lysing erythrocytes by utilizing either monocytes or NK cells, because significant (50%) lysis occurred with effector cell populations magnetically depleted through either FcRII or FcRIII. A major difference between these two types of antibody derivative was their ability to function in the presence of high concentrations of normal human Fc. The lysis mediated by BsAb reactive with FcRI or II was unaffected by the presence of human Fc at 2.5 mg/ml (a concentration comparable with that yielded by IgG in plasma) whereas the BsAb recognizing FcRIII and all the Fc-containing derivatives were completely inhibited.This work has been supported by Tenovus, the Cancer Research Campaign, the Leukaemia Research Fund, Italfarmaco, Milano, Italy and the Imperial Cancer Research Fund 相似文献
7.
A comparison of specific antibodies induced by unfolded actins modified either by oxidation or by arylation of lysine residues was reported. We have focused our work on binding properties with filamentous actin and located its preferential antigenic sites for the anti-arylated-actin antibodies in the C-part of the molecule. An interference of anti-oxidized actin antibodies upon actin polymerisation has also been reported. 相似文献
8.
本文以抗人C_(?)的羊IgG为包被抗体,以HRP-HBs抗体为指示抗体,建立了可检测激活补体类HBsAg循环免疫复合物(HBsAg/C3-CIC)的C_3捕捉法酶联免疫吸附试验。检测了236例六种类型临床诊断为乙型肝炎的病人血清标本,其阳性率分别为:无症状携带者(ASC)12.9%(4/31),急性肝炎(AH)36.7%(22/60),慢性迁延性肝炎(CPH)33.3%(7/21),慢性活动性肝炎(CAH)59.6%(34/57),重型肝炎(SH)77.8%(14/18),肝炎后肝硬化(PLC)67.3%(33/49),阳性率与肝损严重程度明显相关(P<0.01)。认为HBs-Ag/C3-CIC可能在乙型肝炎病毒引起的慢性活动性肝炎、重型肝炎和肝炎后肝硬化的发病过程中起重要作用,并可作为乙型肝炎的诊断、临床分型和预后判断的指标之一。 相似文献
9.
应用SDS-聚丙烯酰胺凝胶电泳及免疫转印技术对流行性出血热患才血清中免疫合物组分进行了分析。流行性出血热循环免疫复合物经SDS-PAGE分离,考马斯亮兰染色,显色主要有7条带,分子量分别为23kD,50kD,52kD,65kD,72kD,80kD及100kD。采用该病毒特异性抗血清、单克隆抗体以及人免疫球蛋白、补体成分抗血清识别,在其特环免疫复合物中可检出特异性病毒抗在及相应的免疫球状蛋白和补体成 相似文献
10.
Peter E. Molloy Barbara M. Graham Pauline M. Cupit Steven D. Grant Andrew J. R. Porter Charles Cunningham 《Molecular biotechnology》1995,4(3):239-245
This work describes protocols for the production of single-chain antibody and T-cell receptor fragments inE. coli. A choice of methods is given for the purification of the recombinant fragments that rely on the use of either immunoaffinity
or metal chelate affinity chromatography. The TCR fragments may have to be denatured and refolded before the fragments attain
their proper conformation. 相似文献