TubulinPolymerizationwithGTP／GMPCPP／Taxol
I. Solutions & SuppliesII. Prepolymerization ClarificationIII. GTP PolymerizationIV. Taxol PolymerizationV. GMPCPP PolymerizationVI. Determining Concentration of GMPCPP/Taxol MTsBack to protocolsI. Solutions & SuppliesBRB80 (1X): 80 mM PIPES, 1 mM MgCl2, 1 mM EGTA, pH 6.8 with KOH (generally made as a 5X stock and stored at 4¡C)100 mM GTP100 mM GMPCPPTaxol: 10 mM stock; 100 µM, 10 µM and 1 µM dilutions (......阅读全文
Tubulin－Polymerization－with－GTP/GMPCPP/Taxol
I. Solutions & SuppliesII. Prepolymerization ClarificationIII. GTP PolymerizationIV. Taxol PolymerizationV. GMPCPP PolymerizationVI. Determining Conce
Recycling－Tubulin
Recycling TubulinWe "recycle" tubulin fractions stored at -80¡C after the PC column and store the recycled tubulin in small aliquots for day-to-day us
Tubulin－Basics
I. Useful Values1 mg/ml tubulin = 10 µM (assuming MW of ab-tubulin heterodimer is 100,000; in reality it is ~110,000 but almost all tubulin labs use t
Tubulin－Basics
I. Useful Values1 mg/ml tubulin = 10 µM (assuming MW of ab-tubulin heterodimer is 100,000; in reality it is ~110,000 but almost all tubulin labs use t
Preparation－of－tubulin
Although many protocols for tubulin preparation are available, the procedure described below is the simplest and highest yielding preparation I have d
Tubulin－Preparat
Materials3 - 5 Fresh Pig Brains1 M GTP1 M Magnesium SulfatePM buffer =100 mM Pipes, pH 6.9 2 mM EGTA 1 mM Magnesium Sulfate2 mM DTTPM-4M Buffer =100 m
Labeling－Tubulin－and－Quantifying－Labeling－Stoichiometry2
II. Labeling ProtocolThe procedure described below can be scaled down if desired. It is essential to perform all steps involving caged dyes under a sa
Immunofluorescent－Localization－of－Tubulin
LEVEL IIMaterialsCoverslip cultures of an appropriate monolayer cell linePhosphate buffered saline (PBS)Acetone/Methanol (absolute) in a 50:50 volume
细胞组分和细胞器——细胞骨架
Fixation and Immunofluorescence of the Cytoskeleton (Mitchison Lab) Recycling Tubulin (Mitchison Lab) Labeling Tubulin and Quantifying Labeling Stoi
Large－Scale－Tubulin－Preparation
Tubulin is purified from bovine/porcine brain by two cycles of polymerization/depolymerization followed by removal of copurifying proteins on a phosph
Preparation－of－tubulin2
DAY 2: Cycling preparation of MT protein.Keep the brains in an evacuated plastic ziplock bag buried in ice from the time of slaughter during transport
porcine－brain－tubulin－prep
Although many protocols for tubulin preparation are available, the procedure described below is the simplest and highest yielding preparation I have d
SDS－Gel－Electrophoresis－of－Tubulin\MAPs
MaterialsStock Acrylamide: (30%T:0.8%C)30% by weight of acrylamide0.8% by weight of N,N'-bis-methylene acrylamideSeparation Gel (Final Concentrati
Large－Scale－Tubulin－Preparation——2
III. Pouring a 1L Phosphocellulose (PC) ColumnResin: Whatman P11 Cellulose Phosphate -- fibrous cation exchanger(1 gram of PC swells to about 4 ml pac
Labeling－Tubulin－and－Quantifying－Labeling－Stoichiometry
Labeling Tubulin and Quantifying Labeling StoichiometryThis is a general procedure for coupling moieties with reactive succinimidyl esters to tubulin.
Isolation－of－Microtubules－(Bovine－Brain)
LEVEL IIMaterialsFreshly removed bovine brain 2Wire sieve (tea strainer)Microtubule buffer (MT buffer)0.1 M MES (2-(N-Morphilino)ethanesulfonic acid)1
Preparation－of－Segmented－and－Polarity－Marked－Microtubules
Segmented and polarity-marked microtubules are very useful for many different types of in vitro assays. Segmented microtubules are microtubules with a
Viscosity－－Polymeriztion－of－Microtubules
LEVEL IIMaterialsTubulin (Brain extract from Exercise 9.4)GTPATPViscometerWaterbath or incubator at 37° CProcedureCompute the amount of GTP (M.W. 523)
Microtubule－(MT)/Organelle－Motility－Assays
Rapidly thaw and immediately place on ice one aliquot each of axonemes, Golgi or ER membranes, 45 uM tubulin, rat liver cytosol, and 20x energy regene
Preparation－of－Segmented－and－Polarity－Marked－Microtubules
Preparation of Segmented and Polarity Marked Microtubules Segmented and polarity-marked microtubules are very useful for many different types of in vi
βTubulin抗体—做出漂亮WB和IHC图片的内参
Tubulin即微管蛋白，是细胞的一种骨架蛋白。Tubulin分为α、β、γ、δ、ε等多种tubulin，其中α-Tubulin和β-Tubulin可以形成异源二聚体，是形成微管的最主要的两种Tubulin。α-tubulin和β-tubulin分子量分别为55kDa和50kDa，其实际检测条带均在
MT－Spindowns－from－Extracts
MT Spindowns from ExtractsArshad DesaiNotes:The key variable in MT spindown experiments is ATP. Under high ATP conditions,conventional MAPs are select
Negative－Stain－Electron－Microscopy－of－Microtubules
Negative staining is a rapid, qualitative method for analyzing microtubule structure at the EM level. Because negative staining involves deposition of
作用于微管系统的药物研究（四）：tubulin－code与药物靶点
有位同行曾经对我说：你在紫杉烷与微管作用及肿瘤耐药方面的研究虽然做得还不错，但无论是靶点还是药物分子都太老了。言下之意这样研究很难继续走下去。考虑到当今时代促使人们去弄潮（也可称之为追逐热点—其利与弊在这里不展开了），而且科研确实也应该不断探索未知领域，不能满足于对已有知识体系的修修补补，在这个
TUBA3C基因编码的功能和结构描述
真核细胞骨架的微管具有多种功能，由α和β微管蛋白的异二聚体组成。编码这些微管成分的基因是微管蛋白超家族的一部分，该家族由六个不同的家族组成。所有真核生物中都有α、β和γ微管蛋白家族的基因。α和β微管蛋白是微管的主要成分，而γ微管蛋白在微管组装成核过程中起着关键作用有多种α和β-微管蛋白基因，在种间和
TUBA3C基因突变因子与药物介绍
真核细胞骨架的微管具有多种功能，由α和β微管蛋白的异二聚体组成。编码这些微管成分的基因是微管蛋白超家族的一部分，该家族由六个不同的家族组成。所有真核生物中都有α、β和γ微管蛋白家族的基因。α和β微管蛋白是微管的主要成分，而γ微管蛋白在微管组装成核过程中起着关键作用有多种α和β-微管蛋白基因，在种间和
PNAS：微管蛋白影响发育的不对称性
机体发育过程中，内脏器官以一种一致性的不对称形式排列——心脏和胃在左边，肝和囊尾在右边，而这一切是如何发生的呢? 美国Tufts大学的生物学家得到了微管蛋白tubulin在许多物种发育早期形成不对称模式的第一手证据，包括植物、线虫、青蛙和人体细胞。文章发表在7月16日Proceedings
TUBA1A基因突变因子与药物介绍
真核细胞骨架的微管具有重要和多样的功能，由α和β微管蛋白的异二聚体组成。编码这些微管成分的基因属于微管蛋白超家族，由六个不同的家族组成。所有真核生物中都有α、β和γ微管蛋白家族的基因。α和β微管蛋白是微管的主要成分，而γ微管蛋白在微管组装成核过程中起着关键作用。有多种α和β-微管蛋白基因，在物种间高
TUBA1A基因编码的功能和结构描述
真核细胞骨架的微管具有重要和多样的功能，由α和β微管蛋白的异二聚体组成。编码这些微管成分的基因属于微管蛋白超家族，由六个不同的家族组成。所有真核生物中都有α、β和γ微管蛋白家族的基因。α和β微管蛋白是微管的主要成分，而γ微管蛋白在微管组装成核过程中起着关键作用。有多种α和β-微管蛋白基因，在物种间高
TTL基因编码的功能和结构描述
ttl是一种参与α-微管蛋白翻译后修饰的胞浆酶（见mim 602529）。聚集微管内的α-微管蛋白随着时间在C末端被去甲状腺激素化微管解体后，TTL恢复酪氨酸残基，从而参与微管蛋白去酪氨酸酶和酪氨酸酶的循环（ERCK等人，2003【PubMed 14571137】。TTL is a cytosoli