Twodimensionalpeptidemapping
This specfic protocol is the latest incarnation of peptide mapping procedures that have been developed here in the TVL/MBVL of the Salk Institute over the last 20 years. Wade Gibson developed the first peptide mapping protocol. This was improved by Karen Beemon and Tony Hunter. Tamara Hurley and Kunxin Luo adapted it to proteins transferred to filters.Procedure:1. Immunoprecipitate ......阅读全文
Two-dimensional-peptide-mapping
This specfic protocol is the latest incarnation of peptide mapping procedures that have been developed here in the TVL/MBVL of the Salk Institute over
peptide-fingerprint-mapping
Polyacrylamide gel electrophoresis is a widely used technique to separate proteins from biological samples. Moreover, the development of two-dimension
Ingel-digestion-of-proteins-for-peptide-fingerprint-mapping
Polyacrylamide gel electrophoresis is a widely used technique to separate proteins from biological samples. Moreover, the development of two-dimension
肽段
· Designing Your Peptide (Genosys)· Handling & Storage of Peptides (Genosys)Two Dimensional Peptide Mapping (Sefton Lab) Synthesizi
Two-Dimensional-Gel-Electrophoretic-Analysis-for-the-Human-Plasma-Proteome
OverviewThis protocol is a detail description of the laboratory procedure in performing 2D gel electrophoresis for illustrating the protein profile of
双向电泳(twodimensional-electrophoresis)完整操作步骤
(一)第一向等电聚焦从冰箱中取-20℃冷冻保存的水化上样缓冲液(I)(不含DTT,不含Bio-Lyte)一小管(1ml/管),置室温溶解。2. 在小管中加入0.01g DTT, Bio-Lyte 4-6、5-7各2.5ml,充分混匀。3. 从小管中取出400ml水化上样缓冲液,加入100ml样品,充
双向电泳(twodimensional-electrophoresis,2DE)2
2.[操作步骤] 1. 将研磨管离心一分钟(不低于12000×g)弃上清。 2. 加入裂解液(200-300ul)充分vortex. 3. 再加入样品鼠脑组织(低于100mg)充分研磨。然后可以再加入裂解液至1ml.. 4. 将组织悬液离心5-10min(高于12000
双向电泳(twodimensional-electrophoresis,2DE)3
2.[操作步骤]1. 将标准品BSA(5mg/ml)先稀释成0.5 mg/ml,2. 按0, 1, 2, 4, 8, 12, 16, 20μl分别加到96孔板中,加DDW补足到20μl。3. 加适当体积样品(4μl)到96孔板的样品孔中,加DDW到20μl。4. 各孔加入200μl G-250染色液
Study-on-a-TwoDimensional-Scanning-MicroMirror-and-Its-Application-...1
Study on a Two-Dimensional Scanning Micro-Mirror and Its Application in a MOEMS Target DetectorChi Zhang,* Zheng You, Hu Huang, and Guanhua LiAuthor i
双向电泳(twodimensional-electrophoresis,2DE)4
由于合成载体两性电解质(synthetic carrier ampholyte SCA)是通过复杂的合成过程得到的,其重复性很难控制,由此不同批次之间会存在很大的变化,同一蛋白质在不同批 图-1. 等电聚焦的“聚焦效应” 次等电聚焦中所出现的位置有所偏差,这样作为双向电泳中的一向时就限
双向电泳(twodimensional-electrophoresis,2DE)6
我们本次实验使用的是银染。银染的方法种类很多,目前有文献报道的就有100多种。但是其准确的染色机制还不是特别的清楚。大致的原理是银离子在碱性pH环境下被还原成金属银,沉淀在蛋白质的表面上而显色。 由于银染的灵敏度很高,可染出胶上低于1 ng/蛋白质点,故广泛的用在2D凝胶分析上。待找到自己感
双向电泳(twodimensional-electrophoresis,2DE)5
六、第二向 SDS-PAGE1.[基本原理]蛋白质在聚丙烯酰胺凝胶中电泳时,它的迁移率取决于它所带净电荷以及分子的大小和形状等因素。如果加入一种试剂消除电荷、形状等因素的影响,使电泳迁移率只取决于分子的大小,就可以用电泳技术测定蛋白质的分子量。1967年,Shapiro等发现在样品介质和聚丙烯酰胺凝
Study-on-a-TwoDimensional-Scanning-MicroMirror-and-Its-Application-...2
In order to realize the decoupling measurement for two deflection angles and obtain the large piezoresistive coefficients for the high measureme
Study-on-a-TwoDimensional-Scanning-MicroMirror-and-Its-Application-...3
2.4. CharacteristicsThe two resonance frequencies of the two-dimensional scanning micro-mirror are 216.8 Hz and 464.8 Hz, respectively, which are
双向电泳(twodimensional-electrophoresis,2DE)1
一、蛋白质组学概论随着人类基因组计划的实施,生命科学步入了后基因组时代,出现了不同于以往经典生物实验科学的全新的研究方式─“生物大科学”。这种生物大科学的核心思想是整体性研究,即以生物体内某类物质为对象进行完整的研究。过去对生命活动的研究仅限于研究细胞内个别的基因或蛋白质,而基因组学和蛋白质组学的目
Study-on-a-TwoDimensional-Scanning-MicroMirror-and-Its-Application-...4
The target ranging is based on the phase-shift laser ranging method. The phase difference between the modulated signal and the reflected signal
Peptide-map-prediction
Peptide map predictionIn identifying peptides from proteins with a known sequence, it is often useful to be able to predict how a peptide will migrate
Peptide-map-prediction
In identifying peptides from proteins with a known sequence, it is often useful to be able to predict how a peptide will migrate during electrophoresi
双向电泳(twodimensional-electrophoresis)操作步骤及相关溶液
实验相关试剂配制1.Bradford 工作液95%乙醇 25ml 先用乙醇溶解考马斯亮兰G250,溶解完后再加磷85%磷酸 52ml 酸,最后超纯水定容至500ml.过滤后置于棕色瓶考马斯亮兰G250 0.035g 外加油皮纸保存(Bradford不稳定,一周内有效)2.裂解液尿素 8M硫脲 2MC
蛋白质双向电泳(twodimensional-electrophoresis)过程与体会2
2.3.2 第二向SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)选用DYY-Ⅲ型(北京六一仪器厂生产)电泳槽(规格为200×200×1mm),分离胶浓度为12%,无浓缩胶。待胶聚合后,将电聚焦后已经平衡的胶条平放于浓缩胶顶端(避免胶条拉直),并用1%琼脂糖(电极缓冲液配制)封胶,特别应注意避免胶条与
蛋白质双向电泳(twodimensional-electrophoresis)过程与体会1
双向电泳IEF (sigma)IEF(not IPG)/SDS-PAGE最简单的装备推荐如下,适合于没多少money做IPG又想做“热”的所谓蛋白质组的,嘻嘻!IEF用Biorad的圆盘电泳槽(不行用国产的吧,不推荐),SDS-PAGE用六一厂的就可以了(ft,六一厂应该给我money吧,给你做广告
蛋白质双向电泳(twodimensional-electrophoresis)过程与体会3
二、一向电泳(13cm的holder)(1)取大约70-100ng的蛋白与溶胀液混合总体积达到250vl(2)将上述溶液加到holder 的两个电极之间。(3)去掉胶条的保护膜,胶面朝下,先将胶条尖端朝胶条槽的尖端方向放入胶条槽中,慢慢下压胶条,并前后移动,避免生成气泡,最后放下胶条平端,使溶液浸湿
氨基酸分离与鉴定——双向层析法(twodimensional-chromatograp...
实验原理纸层析是以滤纸作支持物,用一定的溶剂系统展开,使混合样品达到分离分析的层析方法。其一般操作是将样品溶解在适当溶剂中,点样在滤纸的一端;再选用适当的溶剂系统,从点样的一端通过毛细现象向另一端展开,展开完毕,取出滤纸晾干或烘干,再以适当的显色剂或紫外灯、荧光灯下观察其图谱。样品经展开后某一物质在
二维聚丙烯酰胺凝胶电泳(twodimensional-polyacrylamide-gel-el
二维聚丙烯酰胺凝胶电泳技术结合了等电聚焦技术(根据蛋白质等电点进行分离)以及SDS-聚丙烯酰胺凝胶电泳技术(根据蛋白质的大小进行分离)。这两项技术结合形成的二维电泳是分离分析蛋白质最有效的一种电泳手段。通常第一维电泳是等电聚焦,在细管中(φ1~3 mm)中加入含有两性电解质、8M的脲以及非离子型去污
EDSmapping与EELSmapping的区别
如果是做STEM的EELS mapping时间比edx mapping只长不短。分辨率是eels mapping比edx mapping高
Peptide-competition...
实验概要The peptide competition assay/blocking procedure can be used in western blotting, IHC and ELISA. The peptide competition assay (PCA) is a reco
Preparation-of-Stroma,-Thylakoid-Membrane,-and-Lumen-Fractions-from-...
Preparation of Stroma, Thylakoid Membrane, and Lumen Fractions from Arabidopsis thaliana Chloroplasts for Proteomic AnalysisFor many studies regarding
Comprehensive-identification-of-novel-proteins-and-Nglycosylation-sites一
Comprehensive identification of novel proteins and N-glycosylation sites in royal jellyLan Zhang1,2†, Bin Han1†, Rongli Li1, Xiaoshan Lu1,3, Aiying Ni
Blocking-with-immunizing-peptide-protocol
实验概要The method provides a blocking with immunizing peptide protocol.实验原理Non-specific binding of an antibody to proteins other than the antigen can s
反向微柱的准备Preparation-of-ReversedPhase-Microcolumns
INTRODUCTIONOne versatile strategy for sample cleanup prior to MALDI-MS analysis uses microscale columns designed for direct sample elution onto the M