TubulinBasics
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 this convenient conversion relationship)1 µm of a microtubule has ~1600 tubulin heterodimersExtinction coefficient of tubulin at 280 nm = 115,000 M-1cm-1 or 1.15 (mg/ml)-1 cm-1; assuming that the MW of a tubulin heterodimer is 100,000 daltons. This extinction c......阅读全文
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
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
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
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
Immunofluorescent－Localization－of－Tubulin
LEVEL IIMaterialsCoverslip cultures of an appropriate monolayer cell linePhosphate buffered saline (PBS)Acetone/Methanol (absolute) in a 50:50 volume
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
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
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
Basics－of－Electrochemical－Impedance－Spectroscopy（一）
This tutorial presents an introduction to Electrochemical Impedance Spectroscopy (EIS) theory and has been kept as free from mathematics and ele
Basics－of－Electrochemical－Impedance－Spectroscopy（六）
Purely Capacitive CoatingA metal covered with an undamaged coating generally has a very high impedance. The equivalent circuit for such a situatio
Basics－of－Electrochemical－Impedance－Spectroscopy（九）
Multiple ModelsThe impedance spectrum in Figure 25 shows two clearly defined time constants.Figure 25. Two Time Constant SpectrumThis spectrum can be
Basics－of－Electrochemical－Impedance－Spectroscopy（四）
We will calculate two examples to illustrate a point about combining circuit elements. Suppose we have a 1 Ω and a 4 Ω resistor in series. The i
Basics－of－Electrochemical－Impedance－Spectroscopy（八）
EIS of Coated MetalsThe impedance behavior of a purely capacitive coating was discussed above. Most paint coatings degrade with time, resulting in
Basics－of－Electrochemical－Impedance－Spectroscopy（二）
With Eulers relationship,(6) it is possible to express the impedance as a complex function. The potential is described as,(7) and the current response
Basics－of－Electrochemical－Impedance－Spectroscopy（七）
Mixed Kinetic and Diffusion ControlConsider a cell where semi-infinite diffusion is the rate determining step, with a series solution resistance a
Basics－of－Electrochemical－Impedance－Spectroscopy（三）
In normal EIS practice, a small (1 to 10 mV) AC signal is applied to the cell. With such a small potential signal, the system is pseudo-linear.
Basics－of－Electrochemical－Impedance－Spectroscopy（五）
From this equation the exchange current density can be calculated when Rct is known.DiffusionDiffusion also can create an impedance called a Warburg
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.
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
Tubulin－Polymerization－with－GTP/GMPCPP/Taxol
I. Solutions & SuppliesII. Prepolymerization ClarificationIII. GTP PolymerizationIV. Taxol PolymerizationV. GMPCPP PolymerizationVI. Determining Conce
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－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
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
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