PenicillanBindingProteinAssay青霉素与细胞膜蛋白结合实验
Wash cells with 10 mM Tris pH 8French pressSlow speed spinHigh speed spinResuspend in 10 mM TrisSonicate 2 x 15 sec to remove ß-lacatamasesWash in 10 mM TrisResuspend in 10 mM TrisStore at ?70ºC until needed.Assay:Incubate membranes with ~5 µCi 3H-penicillin at 25ºC for 10 minutesAdd 1000X cold penicillinSDS-PAGEIncubate in 1 M sodium salicylate for 30 minutesDry at 80ºC for 2 hours under vacuumExpose to photogr......阅读全文
Penicillan－Binding－Protein－Assay青霉素与细胞膜蛋白结合实验
Wash cells with 10 mM Tris pH 8French pressSlow speed spinHigh speed spinResuspend in 10 mM TrisSonicate 2 x 15 sec to remove ß-lacatamasesWash in 10
Glycolipid－Binding－Assay
Glycolipid Binding AssaySource: Contributed by Pingsunjim, Paller’s LabAbstract: This protocol can be used for the detection of glycolipids binding to
MinichromosomeMicrotubule－Binding－Assay－微染色体－微管结合实验1
Koshland Lab,Carnegie Institute http://www.ciwemb.edu/labs/koshland/Protocols/MICROTUBULE/mmb.htmlDetermine the OD600 and correlate the cell density f
MinichromosomeMicrotubule－Binding－Assay－微染色体－微管结合实验2
YWB per 10ml5mL 2M Sorbitol (if NZ arrested, add 40uL 1.5mg/mL0.336mL 1M K2HPO4 N2 to 4mL YWB)0.064mL 1M KH2PO44.6mL dH2OYWB, glycerol, PMSF5mL 2M Sor
MINICHROMOSOME－MICROTUBULE－BINDING－ASSAY
Determine the OD600 and correlate the cell density from the chart. Set up four 100mL YPD cultures at the following densities: 0.7x105, 1x105, and 3
Nucleotide－Binding/Hydrolysis－Assay
MaterialsNucleotide mixMotor (50 - 100 µM; purity > 95%)0.5 M Tris-OAc, pH 7.510 mM EGTA10 mM MgCl2DDWSephadex G-50 Medium column (0.8 cm in x 20 cm)C
Bradford－protein－assay
Bradford protein assayConsiderations for useThe Bradford assay is very fast and uses about the same amount of protein as the Lowry assay. It is fairly
LOWRY－PROTEIN－ASSAY
The Lowry procedure is one of the most venerable and widely-used protein assays, being first described in 1951 [Lowry et al., J. Biol. Chem. 193: 265-
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAYMATERIALSBiuret ReagentBovine serum albumin (BSA)Spectrophotometer and tubesPROCEDUREPrepare standard dilutions of BSA containing
Protein－Assay－(Spectrophotometer)
Protein Assay (Spectrophotometer)Use BSA (bovine serum albumin) 1mg/ml stock solution (1ml Eppendorf tubes) for standard curve.Place 0, 2, 5, 10, 15,
MINICHROMOSOME－MICROTUBULE－BINDING－ASSAY2
HYBRIDIZATION.Prehybridize blot at 65oC for ~3h in Church buffer containing 0.5mg/ml denature salmon sperm DNA (usually 14ml Church buffer plus 0.7ml
Bradford－Protein－Concentration－Assay
Bradford Protein Concentration Assayversion 01/07/2001Abbreviations:mcg = microgramsmcL = microlitersBSA = bovine serum albuminO.D. = optical densityd
Biorad－Protein－Assay:－Bradford
Biorad Protein Assay: BradfordStandards: 1 mg/ml BSA stock- dilute 1:10 to get 0.1 mg/ml BSAAdd To get H-2O20 µl 2 µg/ml 780 µl40 µl 4 µg/ml 760 µl60
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
Angiotensin－Protein－Kinase－Assay
James Hardwick's angiotensin assay protocolThis specific procedure was developed to assay the activity of the Lck kinase expressed from a retrovir
Transcription,－Translation－of－S35Radiolabelled－Protein－and－Binding－to－GST
Prepare the template by linearizing 25ug plasmid DNA at the 3'' end of the insert. Phenol / chloroform extract, ethanol / NaCl precipitate and
Bradford法蛋白定量（Bradford－Protein－Assay－）
Bradford Assay is a rapid and accurate method commonly used to determine the total protein concentration of a sample. The assay is based on the observ
Use－of－the－Bradford－Protein－Assay－in－a－Microtiter－Plate－Format
Introduction The Bradford protein assay is a simple procedure for determination of protein concentrations in solutions that depends upon the change in
生物素与细胞表面的膜蛋白是怎样结合的
通过生物素化标记分析细胞膜亚蛋白质组Analysis of Cell Membrane Subproteome by Biotinylation of Proteins添加成功！您可以在“我的服务”中查看您添加的引用通知列表，并且配置获取通知的方式。关闭细胞膜在许多基本生物学作用过程中扮演着重要角色
KinExA分子和细胞相互作用仪与SPR的技术对比（二）
四、案例案例一：完整细胞的相互作用检测：单克隆抗体XMetA是胰岛素受体（IR）变构部分的激动剂，其激活代谢Akt激酶信号通路，而对有丝分裂胞外信号调节激酶（ERK）信号通路几乎没有影响。为了研究这种选择性信号通路的性质，作者验证了XMetA对CHO细胞中IR，Akt和ERK的特异性磷酸化和活化的影
肿瘤细胞侵袭实验（Tumour－Invasion－Assay）
实验方法原理 Matrigel 是从小鼠EHS肉瘤中提取的基质成分，含有LN、IV型胶原、接触蛋白和肝素硫酸多糖，铺在无聚乙烯吡硌烷酮的聚碳酸酯滤膜上，能在DMEM培养基中重建形成膜结构，这种膜结构与天然基质膜结构极为相似。 滤膜孔径一般为8um，而且膜孔都被Matrigel覆盖，细胞不能自由穿过，
凝胶迁移滞后实验基本原理
凝胶迁移滞后实验（electrophoretic mobility shift assays，EMSA）是近年发展起来的研究核酸与蛋白质相互作用简单、快速、敏感的方法。目前已经成为转录因子研究的经典方法。其基本原理是蛋白质可以与末端标记的核酸探针结合，电泳时这种复合物比无蛋白结合的探针在凝胶中泳动的
条带转移(Band－Shift)
Or gel mobility shift assay, gel shift assay, gel retardation, electrophoretic mobility shift assay (EMSA) EMSA Using Oligos (Mike A. Dyer)Anneal two
Gel－Shift－Assay－Systems
ProtocolsDownloadprotocol183kbpdf?Abstract for Gel Shift Assay SystemsThe gel shift, or electrophoretic mobility shift, assay provides a simple and ra
染色质蛋白非组蛋白的介绍
非组蛋白主要是指与特异DNA序列相结合的蛋白质,所以又称序列特异性DNA结合蛋白（sequence specific DNA binding protein)。利用凝胶延滞实验（gel retardation assay）,可以在细胞抽提物中进行检测。首先制备一段带有放射性标记的已知特异序列的D
非组蛋白的概念和检测方法
非组蛋白主要是指与特异DNA序列相结合的蛋白质，所以又称序列特异性DNA结合蛋白（sequence specific DNA binding protein)。利用凝胶延滞实验（gel retardation assay）,可以在细胞抽提物中进行检测。首先制备一段带有放射性标记的已知特异序列的DNA
简述青霉素结合蛋白的分类
按照细菌对各种β-内酰胺类抗生素的敏感度，可以将PBPs大致分为两类: ①对青霉素敏感度稍差，但对大多数头孢菌素敏感的PBPs:分子质量较低(Mr24.8～34.7ku)，一般是D，D-肽酶，作用于肽或羧肽羧基供体相似物。如金黄色葡萄球菌的PBP4，大肠埃希氏菌的PBP5、PBP6； ②一般
青霉素结合蛋白的结构介绍
以前是通过细菌提取的羧肽酶的青霉素-肽衍生物的氨基酸序列分析弄清了数种PBPs的活性中心，结果显示细菌PBP活性中心常是丝氨酸，β-内酰胺类抗生素正是通过其β-内酰胺环中的羧基和细菌相应的PBPs的丝氨酸的羟基共价结合成丝氨酸酯起作用。另外也有研究表明可能存在半胱氨酸-巯基为活性中心的羧肽酶。后
青霉素结合蛋白的基本介绍
青霉素结合蛋白(PBPs)是位于细菌细胞膜上的一些膜蛋白,最初发现时因为能和青霉素共价结合而得名。1972年Suginaka等第一次报道了青霉素结合蛋白(PBPs),他们用放射性同位素标记的青霉素标出了细菌表面的PBPs。后来研究发现PBPs也能与非β-内酰胺类抗生素结合。另外非细菌如螺旋体也可
Chemotaxis－Assay趋化性实验
Springer Lab，The CBR Institute for Biomedical Research, Inc. Department of Pathology Harvard Medical Schoolhttp://cbr.med.harvard.edu/investigators/sp