ImmunofluorescentLocalizationofTubulin
LEVEL IIMaterialsCoverslip cultures of an appropriate monolayer cell linePhosphate buffered saline (PBS)Acetone/Methanol (absolute) in a 50:50 volume mixtureRabbit anti-tubulin (or other primary antibody to tubulin)FITC-labeled goat anti-rabbit (or secondary antibody to match the primary)Fluorescence Mounting MediaFluorescent Microscope equipped with 490 nm excitation filter and 515 nmbarrier filterKodachrome film or......阅读全文
Immunofluorescent－Localization－of－Tubulin
LEVEL IIMaterialsCoverslip cultures of an appropriate monolayer cell linePhosphate buffered saline (PBS)Acetone/Methanol (absolute) in a 50:50 volume
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
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－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
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
Studying－Arabidopsis－Envelope－Protein－Localization－and－Topology－Using－...
Chloroplasts are metabolically important organelles that perform many essential functions within plant cells. The chloroplasts can be subdivided i
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
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.
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
Tubulin－Polymerization－with－GTP/GMPCPP/Taxol
I. Solutions & SuppliesII. Prepolymerization ClarificationIII. GTP PolymerizationIV. Taxol PolymerizationV. GMPCPP PolymerizationVI. Determining Conce
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
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
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
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
Fluorescence－Procedures－fortheActin－andTubulin－Cytoskeleton－in－Fixed－Cells2
Formaldehyde FixationFix in 4% formaldehyde (16% stock EM grade) in CBS for 20 minutesRinse in TBSPermeabilize as for methanol fixationProcede as for
Fluorescence－Procedures－forthe－ActinandTubulin－Cytoskeleton－inFixed－Cells2
Actin CytoskeletonMethanol fixationFix in -20oC methanol for 1-2.5 minutesRinse in TBSPermeabilize in TBS-0.5% TX for 10 minutesRinse in TBS-0.1% TX (
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
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
Fluorescence－Procedures－forthe－Actin－andTubulin－Cytoskeleton－in－Fixed－Cells
Fluorescence Procedures for the Actin and Tubulin Cytoskeleton in Fixed CellsActin: Louise CramerTubulin: Arshad DesaiGeneral StrategyWe typically wor
Immunofluorescent－Staining－of－Drosophila－Larval－Brain－Tissue
INTRODUCTIONThe Drosophila larval brain is a well-established model for investigating the role of stem cells in development. Neuroblasts (neural stem
Fluorescence－Procedures－forthe－ActinandTubulin－Cytoskeleton－inFixed－Cells1
General StrategyWe typically work with tissue culture, primary mammalian cells, and cell extracts, but the protocols can be adapted to other systems,
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
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
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
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