一种简便的考马斯亮蓝G250蛋白质染色方法

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.     

改良的蛋白质电泳考马斯亮蓝染色方法研究

旨在建立简便、经济、灵敏以及质谱兼容的蛋白质聚丙烯酰胺凝胶电泳染色法,在胶体考染(Blue Silver)配方(0.12%的考马斯亮蓝G-250,10%的磷酸,10%的硫酸铵,和20%的甲醇)的基础上,通过优化凝胶固定时间与染色液中各物质用量,得到改良的染色液配方(0.1%的考马斯亮蓝G-250或R-250,5%的磷酸,5%的硫酸铵,10%的甲醇)及简便的操作步骤。试验对比结果显示,改良方法一与胶体考染相比仍保持了较高的染色灵敏度以及较好的质谱兼容性,且有机试剂使用量更少,染色深度与蛋白质量的线性关系也更好。     

荧光素钠和考马斯亮蓝应用于小麦白粉病菌染色效果比较

比较了荧光素钠和考马斯亮蓝应用于小麦白粉菌染色的效果。荧光素钠法中样品处理只需20min左右,具有直接,快速的特点;荧光指示剂对病菌分生孢子萌发及菌丝生长无抑制作用,主要沉集于活菌体的隔膜和细胞质部位,使病菌产生明显的亮绿荧光和清晰的细胞轮廓,亮绿荧光衰退期为7min;借助荧光显微镜可以观察病菌在小麦叶表的发展过程,区别活菌体和失活菌体。考马斯亮蓝法包括传统的组织学染色步骤,经过改进后的样品处理过程需要40min左右;染后使寄主组织呈现淡蓝色,病菌菌体染成深蓝色;该方法可以观察病菌在小麦叶表和被侵染细胞内部发育形成的结构,包括孢子发育形成的初生芽管,附着胞芽管,成熟附着胞以及在寄主细胞内形成的初生吸器原体,成熟的指状体吸器和次生吸器。     

蛋白质组学实验中聚丙烯酰胺凝胶的考马斯亮蓝染色

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。     

粗毛栓菌产漆酶对考马斯亮蓝的脱色降解

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。     

一种简易的与考马斯亮蓝相结合的低背景银染法

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     

考马斯亮蓝显色液组分对蛋白质测定的影响

考马斯亮蓝（CBB）显色法测定蛋白质含量的主要缺点之一是线性关系差．通过研究显色液组分对线性关系的影响,发现显色液H⁺浓度是影响线性关系的主要因素,并提出了一个新的显色液配方来改善考马斯亮蓝蛋白质测定法的线性关系．     

考马斯亮蓝染色双向电泳凝胶胶内酶切方法的改进

介绍一种简便高效的从考马斯亮蓝染色的双向电泳凝胶切取蛋白质点,通过胶内酶切制备基质辅助激光解吸电离飞行时间质谱(MALDI—TOF-MS)样品的方法。该方法操作简便,样品的质谱鉴定结果经网上数据库检索检出率可高达80％以上,适于我国目前小规模蛋白质组学研究的国情,便于推广应用。     

考马斯亮蓝法检测重组碱性成纤维细胞生长因子

目的：建立考马斯亮蓝法（CS）检测重组碱性成纤维细胞生长因子（r-bFGF)含量。方法：以牛血清白蛋白（BSA）为标准液,考马斯亮蓝（CBG－250）为染色剂,检测595nm吸光度并绘制标准曲线找出回归方程,对照r-bFGF样品计算含量。结果：此法敏感性高,可测出μg水平蛋白,标本量只需50－100μl,染料与蛋白的结合反应快,只需2min即可比色,显色稳定,可维持1h,重现性好,变异系数在5％以内。结论：此法简便有效、灵敏性高,能快速检测r-bFGF含量。     

Phytoplankton Staining Using Coomassie Brilliant Blue G

A method is introduced to a) distinguish phytoplankton organisms from organic matter and b) ease its determination by staining protein structures with Coomassie Brilliant Blue G.     

Environmentally safe removal/disposal of Coomassie Brilliant Blue from gel destain and used gel stain

Gel destaining following Coomassie Brilliant Blue (CBB) staining involves the use of toxic reagents. Here we demonstrate the efficacy of various paper adsorbents in adsorbing CBB. Kimwipes adsorbed the best, followed by Teri towels, multifold towels, and Whatman numbers 1 and 3 filter papers. Three Kimwipes completely adsorbed the dye released from a CBB-stained mini-gel. Nonradioactive destain solution can, therefore, be recycled for destaining CBB-stained gels. Stain removal with Kimwipes helps in reducing destain use and in reducing organic liquid waste, and it is 7.5-fold cheaper compared with an available method for CBB disposal. Following this, we determined the suitability of this procedure to remove the dye from a used CBB staining solution awaiting proper disposal by our Institutional Safety Office. The dye from a 0.05% CBB staining solution could be removed in 5 to 10 min using 75 Kimwipes. The CBB-adsorbed Kimwipes did not release the stain when squeezed dry even after incubation in various salts over 1 week and in water for 5 weeks. The CBB removed allows its easy disposal as solid waste and will not leach out from solid landfills. Thus, stain removal with Kimwipes helps in disposing CBB in an environmentally friendly manner and allows recycling of destaining solution.     

考马斯亮蓝与牛血清白蛋白相互作用机理的研究

利用光谱探针技术研究在酸性溶液中考马斯亮蓝G-250(Coomassie brilliant blue G-250,CBBG)与牛血清白蛋白(bovine serum albumin,BSA)相互作用机理,考察了不同实验条件对CBBG-BSA复合物吸收光谱的影响。实验结果表明:CBBG与BSA相互作用产生光谱蓝移主要是由CBBG与BSA间的疏水相互作用引起,而静电作用则是形成CBBG-BSA蓝色复合物的必要条件。同时,CBBG聚集体的聚集程度是影响CBBG-BSA蓝色复合物形成的重要因素。     

A modified Coomassie Brilliant Blue staining method at nanogram sensitivity compatible with proteomic analysis

A more sensitive and convenient Coomassie Brilliant Blue (CBB) staining method for visualizing proteins was developed. Compared with the modifications include the supplement of 10% (v/v) methanol into the fixing solution, an increase of an additional sensitization step and CBB raised from 0.1 to 0.125%. The improved method can detect proteins at nanogram level. The improved method is more sensitive than Blue Silver and more convenient than the Silver protocol. Mass spectrometry results confirmed that it is suitable for subsequent proteomic research.     

Standardization of Staining in Giycosaminoglycan Histochemistry: Alcian Blue, its Analogues, and Diamine Methods

Glycosaminoglycans are identified in tissue sections by various histochemical techniques including staining with alcian blue and its analogues, such as cuprolinic blue and cupromeronic blue, or with high and low iron diamine methods. The variation in staining results is particularly confusing in the case of alcian blue, where not only are several different brands of alcian blue available but also several different staining protocols are used. If the results obtained by these techniques are compared, they often do not match. We have developed a dot blot technique for quality control of glycosaminoglycan histochemistry to standardize the staining protocols. This staining technique enables his-tochemists to test particular batches of alcian blue or its analogues for selective glycosaminoglycan staining, thus improving control of histochemical results. The results obtained using the dot blot assay indicate that it is necessary to test each batch of dye individually to obtain valid results in glycosaminoglycan histochemistry.     

Enhanced thiosulfate-silver staining of proteins in gels using Coomassie Blue and chromium

Potassium chromium sulfate, a new sensitivity enhancer for silver staining of proteins in gels, enhanced the sensitivity of the thiosulfate-silver staining method. The sensitivity could be further improved when potassium chromium sulfate was used in association with another sensitivity enhancer, Coomassie Brilliant Blue R-250 (CBB R-250). The sensitivity of the CBB-chromium modified method to strongly basic proteins such as ribosomal proteins was about 20-fold over that of the published method. This novel method has direct applicability for 2-D gel electrophoresis used in proteomics.     

Combined Lectin Binding and PAS/Alcian Blue Staining in Glycol Methacrylate Sections

Evaluation of cryofixation and paraffin and glycol methacrylate embedding showed that lectin binding was essentially independent of the embedding medium. Fluorescence intensity increased in the following order: glycol methacrylate, paraffin and cryostat sections, The optical resolution increased in the reverse order. Semi-thin glycol methacrylate sections provided satisfactory fluorescence intensities and the best resolution of all embedding techniques applied. Furthermore the lectin treated sections can be stained further using routine histological or specific histochemical methods. The potassium hy-droxide/alcian blue/periodic acid-phenylhydra-zine-Schiff method was used successfully to demonstrate sulfated and nonsulfated sialomucins. Lectins combined with mucin histochemistry allowed visualization of specific sugar residues in the same glycol methacrylate plastic section.     

A Proposed Structure of Modified Nucleosides Expected to Have High Antiviral Activity and Low Toxicity

This article describes modified nucleosides with deactivated 3′-OH and modified at two or more positions. These are expected to have high antiviral activity as well as have low toxicity.     

Activation of Mouse Oocytes, Fertilization and Development of Mouse Embryos in Vitro after Staining with Trypan Blue or Fluorescein Diacetate

The influence of vital staining with trypan blue or fluorescein diacetate on the fertilization of mouse oocytes and the developmental potential of mouse embryos was assessed. Neither stain induced spontaneous activation in mouse oocytes, nor did they impair the in vitro development and implantation of mouse zygotes, two-cell embryos, stressed morulae or blastocysts. However, fertilization and subsequent development of mouse oocytes have been shown to be reduced by vital staining.     

Biomimetic Hydrophobic Surfaces with Low or High Adhesion Based on Poly(vinyl alcohol) and SiO2 Nanoparticles

Superhydrophobic surfaces are often found in nature,such as plant leaves and insect wings.Inspired by superhydrophobic phenomenon of the rose petals and the lotus leaves,biomimetic hydrophobic surfaces with high or low adhesion were prepared with a facile drop-coating approach in this paper.Poly(vinyl alcohol) (PVA) was used as adhesive and SiO2 nanoparticles were used to fabricate surface micro-structure.Stearic acid or dodecafluoroheptyl-propyl-trimethoxysilane (DFTMS) were used as low surface energy materials to modify the prepared PVA/SiO2 coating surfaces.The effects of size of SiO2 nanoparticles,concentration of SiO2 nanoparticle suspensions and the modifications on the wettability of the surface were investigated.The morphology of the PVA/SiO2 coating surfaces was observed by using scanning electron microscope.Water contact angle of the obtained superhydrophilic surface could reach to 3°.Stearic acid modified PVA/SiO2 coating surfaces showed hydrophobicity with high adhesion.By mixing the SiO2 nanoparticles with sizes of 40 nm and 200 nm and modifying with DFTMS,water contact angle of the obtained coating surface could be up to 155° and slide angle was only 5°.This work provides a facile and useful method to control surface wettability through changing the roughness and chemical composition of a surface.