共查询到17条相似文献,搜索用时 78 毫秒
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一种简便的考马斯亮蓝G250蛋白质染色方法 总被引:9,自引:0,他引:9
介绍一种快速、简便、几乎无背景的考马斯亮蓝G250(CBB G250)染色方法.该方法所用试剂仅为稀盐酸和CBB G250, CBB G250的工作浓度为0.0015%,灵敏度达0.02 μg/带, 染色2 h达70%,4 h以上或染色过夜即可充分染色.与以往的考马斯亮蓝染色方法相比,该方法有经济方便、灵敏度高、几乎无背景等优点,便于推广应用. 相似文献
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1972年Kerenyi建立的凝胶电泳的银染色法由于灵敏度高而被广泛采用。此法创建初期常产生较深的背景染色,以致影响弱带的显示,后来几经改进,特别是Blum等人利用硫代硫酸钠的化学特性,发展了一种高灵敏度的低背景银染方法,使此法愈 相似文献
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蛋白质组学实验中聚丙烯酰胺凝胶的考马斯亮蓝染色 总被引:1,自引:0,他引:1
在蛋白质组学实验中,蛋白质经聚丙烯酰胺凝胶电泳后的染色是一个十分重要的环节。鉴于其良好的质谱兼容性和操作上的便捷,考马斯亮蓝染色是目前众多实验室中最常用的染色方法。本文介绍了四种各具特色的考马斯亮蓝染色方法。 相似文献
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粗毛栓菌产漆酶对考马斯亮蓝的脱色降解 总被引:1,自引:0,他引:1
粗毛栓菌粗酶液经聚丙烯酰胺凝胶电泳,在CBBG-250染色后,漆酶带处有一透明圈,经纯化漆酶和CBBG-250溶液直接作用以及菌体的培养结果证实,漆酶对CBBG-250具有一定的脱色降解作用,在漆酶活力达118u/ml时,CBBG-250溶液在595nm波长的光吸收60min内下降了32.4%。该粗毛栓菌产漆酶对工业染料废水也具有一定的降解脱色作用。 相似文献
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一种改良的肌细胞骨架染色方法 总被引:5,自引:0,他引:5
为了观察肌细胞骨架,对传统考马斯亮蓝染色法进行改良,并与免疫荧光染色法进行了比较。培养的血管平滑肌细胞先用多聚甲醛预固定后再进行考马斯亮蓝染色,可使细胞骨架非常清晰的显色,解决了传统考马斯亮蓝染色易使肌细胞变形、脱片的问题,其效果与免疫荧光染色相近。因此,多聚甲醛预固定.考马斯亮蓝染色法是一种适于肌细胞骨架染色的简便方法。 相似文献
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本文涉及了双向电泳过程中的染色方法,即先用考马斯亮蓝染色,将胶上可见蛋白切下再银染的方法。这种方法可最大限度的减少胶中蛋白质点的损失,不仅避免了单一用考马斯亮蓝染色由于灵敏度不高而导致的低丰度蛋白的损失,也避免了单一用银染而使高丰度的蛋白因染色过度导致的损失。同时两种传统的染色方法结合完美,形成的新方法经济实用。 相似文献
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蛋白质定量方法的进展 总被引:23,自引:1,他引:23
本文在简要比较常用蛋白质定量方法的基础上,结合自己的工作,选择性地叙述了Lowry法、考马斯亮蓝G-250染料测定蛋白质的改进法。还介绍了银染色定量法和4-甲酸喹啉测定蛋白质含量的新方法。 相似文献
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用一种改进的电泳方法测定抗冻蛋白的分子量 总被引:14,自引:2,他引:14
针对抗冻蛋白分子量较小的特点,该文采用了一种改进的Tricine-SDS-PAGE法测定其分子量。采用三层胶系统并在电极缓冲液中以Tricine代替Glycine。 相似文献
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Secretions from amphids, phasmids, and excretory system were stained by incubating nematodes in 0.1% coomassie brilliant blue G-250 in 40% aqueous methanol containing 10% acetic acid on slides with coverslips sealed with nail polish or Zut. Nematodes incubated in this staining solution usually produced copious amounts of secretions from their amphids and excretory pore. Phasmids also stained dark blue, enabling them to be easily observed. Other biological dyes stained these secretions or were useful for differentiating specific morphological features of nematodes. 相似文献
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Nine different polysaccharides were screened for conjugation with laccase and evaluated for pH and thermal stability. All the polysaccharides decreased the thermal and pH stability of laccase at 50 °C and 60 °C, where conjugation with gum Arabic showing the most pronounced effect. Thermal instability of gum Arabic conjugated laccase was affirmed by differential scanning calorimeter while the structural changes in the conjugated laccase responsible for thermal instability was analysed by fluorescence spectrophotometer. The gum Arabic conjugated laccase showed an unusually high tolerance to sodium chloride, thermal instability and lower stability in alkaline conditions. Gum Arabic conjugated laccase was found to decolorize Remazol brilliant blue R in the textile effluent at a slower rate without any microbial growth which was unlike that observed in effluent treated with free laccase. Further, effluent treated with conjugated laccase enabled its reuse as liquor for the dyeing to get desired shade. 相似文献
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Proteins stained with Coomassie brilliant blue on polyacrylamide gels were digested with lysylendopeptidase in the presence of sodium dodecyl sulfate. Peptide production was similar to that under ordinary conditions of digestion. Peptides were recovered easily and efficiently from the gel pieces and separated by HPLC. The present method for preparation of peptides from proteins separated by sodium dodecyl sulfate gel electrophoresis is quite simple and can be used for sequence analysis of proteins in general at the subnanomolar level. 相似文献
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《Expert review of proteomics》2013,10(6):627-634
Gel-based proteomics are the most useful method for protein separation, even when compared with gel-free proteomics. Proteomic analysis by 2D gel electrophoresis (2-DE) with immobilized pH gradients is in turn the best approach to large-scale protein-expression screening. Spots visualization is pivotal for protein identification by mass spectrometry. Commonly used staining methods with excellent mass spectrometry compatibility are coomassie brilliant blue (CBB) or fluorescent dyes. In this study, an implementation of ‘blue silver’ colloidal CBB staining, characterized by high sensitivity and immediate low background, is discussed. The sensitivity of classical, colloidal and ‘blue silver’ CBB staining methods was compared on monodimensional and 2-DE gels. The implementation of the ‘blue silver’ method performs better, provided the physical state of the micelles is respected. An example of a 2-DE of human urine treated with combinatorial peptide ligand libraries demonstrates that implemented ‘blue silver’ can evidence the complexity of the sample. 相似文献
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A micromethod for the quantitation of cellular proteins in Percoll with the Coomassie brilliant blue dye-binding assay 总被引:11,自引:0,他引:11
A simple procedure for the determination of cellular proteins in Percoll-containing samples is described. Percoll precipitated when particulate proteins were solubilized by dilution of the samples in a NaOH-Triton X-100 mixture. After centrifugation at high speed (12,000g), the supernatant was assayed for proteins with the Coomassie brilliant blue dye-binding assay. With an automatic spectrophotometer, 50-microliter aliquots gave a linear response between 0 and 3 micrograms of bovine serum albumin. After a fivefold dilution in the alkali-detergent mixture, proteins in samples containing up to at least 60% Percoll can be accurately quantitated on a standard curve prepared in the absence of Percoll. Because the sensitivity of the assay was better than 100 ng, the procedure outlined in this paper can also be used as a general protein micromethod. 相似文献
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Alan N. Moshell Robert E. Tarone Stanley A. Newfield Alan D. Andrews Jay H. Robbins 《In vitro cellular & developmental biology. Plant》1981,17(4):299-307
Summary A simple, rapid and reproducible test has been developed to measure the viability of cells after irradiation with ultraviolet
light (UV). Epstein-Barr virus-transformed lymphoid lines, derived from patients with xeroderma pigmentosum (XP), were irradiated
with UV, and the post-UV viability of the lymphoid lines was determined by the trypan blue dye exclusion method. The relative
post-UV survival of the patients' lymphoid lines was similar to the relative post-UV survival of the patients' fibroblast
strains. 相似文献