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A background-free, fast protein staining method in polyacrylamide gel electrophoresis using an acidic dye, zincon (ZC) and a basic dye, ethyl violet (EV) is described. It is based on the counterion dye staining technique that employs two oppositely charged dyes to form an ion-pair complex in staining solution. The selective binding of free dye molecules to proteins in acidic solution produces bluish violet-colored bands. It is a rapid and end-point staining procedure, involving only fixing and staining steps that are completed in 1-1.5 h. The detection limit of this method is 8-15 ng of protein that is comparable to the sensitivity of the colloidal Coomassie Brilliant Blue G (CBBG) stain. Due to its sensitivity and speed, this stain may be more practical than any other dye-based stains for routine laboratory purposes. 相似文献
3.
Yun Khoon Liew Vasanthakumari Neela Rukman Awang Hamat Syafinaz Amin Nordin Pei Pei Chong 《Electrophoresis》2013,34(3):397-400
The typical concentration of protein loaded varies from 0.13 to 1.40 μg/μL for a classical silver staining method in 2DE gel. Here, we present a simple modified classical silver staining method by modifying the silver impregnation and development reaction steps. This modified method detects the protein spots at extremely low loaded concentrations, ranging from 0.0048 to 0.0480 μg/μL. We recommend this modified silver staining as an excellent method for the limited biological samples used for silver‐stained 2DE analysis. Altogether, the protocol takes close to two days from first dimension separation to second dimension separation, followed by silver staining, scanning, and analysis. 相似文献
4.
Guo‐Ying Hong Zhong‐Xin Zhu Yuan‐Meng Duan Xuan Zhou Li‐Tai Jin 《Electrophoresis》2013,34(22-23):3171-3179
As a noncovalent fluorescence probe, in this study, salicylaldehyde azine (SA) was introduced as a sensitive fluorescence‐based dye for detecting proteins both in 1D and 2D polyacrylamide electrophoresis gels. Down to 0.2 ng of single protein band could be detected within 1 h, which is similar to that of glutaraldehyde‐silver stain, but approximately four times higher than that of SYPRO Ruby fluorescent stain. Furthermore, comparative analysis of the MS compatibility of SA stain with SYPRO Ruby stain indicated that SA stain is compatible with the downstream of protein identification by LC‐MS/MS. Additionally, the probable mechanism of the SA stain was investigated by molecular docking. The results demonstrated that the interaction between SA and protein was mainly contributed by hydrogen bonding and hydrophobic forces. 相似文献
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超低间隙钛合金低温动态力学性能 总被引:1,自引:0,他引:1
本文利用实验应变模态分析方法,在低温条件下(液氦到室温)研究了航天材料超低间隙钛合金TA7-D试样的动态力学性能,使用时域系统识别技术分析了材料在不同温度点的模态参数,并根据基频给出了其动态模量随温度的变化。 相似文献
7.
细胞凋亡过程中细胞骨架改变的激光共聚焦显微镜观察 总被引:1,自引:0,他引:1
目的:了解细胞凋亡过程中细胞骨架的形态学改变。方法:应用高浓度全反式维甲酸诱导卵巢癌细胞凋亡,激光共聚焦显微镜观察经荧光染色的细胞微管蛋白的形态学变化。结果:凋亡早期,微管蛋白在荧光相由正常按一定方向分布的丝网状结构变成杂乱分布的网状结构,晚期呈细颗粒状分布,荧光强度随凋亡发展而逐渐变弱。讨论:在凋亡过程中,细胞骨架随凋亡发展出现相应的形态学改变,凋亡早期,细胞骨架在分布上出现变化,伴随细胞形态出现变化,凋亡晚期,细胞骨架蛋白发生降解,此时的细胞形态改变是不可逆的。 相似文献
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H. Aoki S. Kosakabe M. Inumaru A. Kuboki S. Ohira M. Kodama 《Journal of Thermal Analysis and Calorimetry》2008,92(2):443-449
D-erythro sphingomyelines (SM) having a defined acyl chain were synthesized with sphingosylphosphorylcholine as a starting material,
and both a structural property and its relating phase transition phenomenon were compared between a symmetric chain length
SM (palmitoyl-SM: C16-SM) and asymmetric chain length SMs (behenoyl-SM: C22-SM, lignoceryl-SM: C24-SM). Furthermore, effect
of increasing a content of asymmetric chain SMs in the mixture systems of C22-SM/C16-SM, and C24-SM/C16-SM was investigated.
The present calorimetric and electron microscopic studies revealed that (1) The main transition enthalpy is smaller for the
asymmetric chain SMs than for the symmetric chain SM by about 3 kJ mol−1, although the acyl chain length is longer for the former than for latter; (2) Relatively small size vesicles (100∼200 nm
diameters) surrounded by one or more lamellae are observed for the asymmetric chain SMs, in contrast to large multilamellar
vesicles (1500∼2500 nm diameters) having at least fifteen stained lamellae for the symmetric chain SM and (3) The coexisting
asymmetric chain SMs cause the decrease in size and multiplicity for the MLV of the symmetric chain SM, simultaneously with
a decrease in the main transition enthalpy. 相似文献
10.