SteadyStateATPaseAssaysCoupledEnzymeSystem
MaterialsTubulin (>5 mg/mL)100 mM Mg·GTP4 mM Taxol in DMSOPM =100 mM PIPES pH 6.82 mM EGTA1 mM Mg2SO4Motor protein (>95% purity; 15-20 µM)Cuvettes (200 µL volume, 10 mm path length)0.5 M Tris-OAc, pH 7.510 mM MgCl210 mM DTTDDW10 mM Mg·ATP30 mM PEP6 mM NADH (Make fresh in 10 mM Tris-OAc pH 7.5, 4.26 mg/mL = 6 mM)PK/LDH (Sigma #P-0294, PK + LDH from Rabbit Muscle in 50% glycerol)Procedure1.Assemble microtubu......阅读全文
Steady－State－ATPase－Assays－Coupled－Enzyme－System
MaterialsTubulin (>5 mg/mL)100 mM Mg·GTP4 mM Taxol in DMSOPM =100 mM PIPES pH 6.82 mM EGTA1 mM Mg2SO4Motor protein (>95% purity; 15-20 µM)Cuvettes (20
ATPase－Assays－with－32PATP
MaterialsPurified Motor Protein, 20-80 µMNucleotide Mix =50 mM Mg·ATP gamma-32P-ATP to give 5 000 - 10 000 cpm/nmol 10 mM HEPES, pH 7.2 1 mM EGTA 1 mM
Nuclear－RunOn－Transcription－Assays
Nuclear “run-on” (or “run-off”) transcription assays have been used to obtain quantitative information about the relative rates of transcription o
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.
Recommended－Experiments－with－Isolated－Mitochondria2
2. Respiratory control on glutamate and inhibition of electron transportAdd 30-40 µl mitochondria, obtain a steady state. We use a larger volume tha
Recommended－Experiments－with－Isolated－Mitochondria
In our teaching lab we encourage students to work with each other and to share insight, experience, and even experimental results. To facilitate such
线粒体荧光探针大全：TMRM，Mitotracker，JC1（5）
Monoclonal Antibodies Specific for Proteins in the Oxidative Phosphorylation SystemOxidative phosphorylation (OxPhos) activity occurs in the mitochond
Invivo－evaluation－ofdrug－lead－candidates－by－intravenous－continuous－infusion
In vivo evaluation of drug lead candidates by intravenous continuous infusionSang Ho LeeWesley ShoopBruce MichaelThomas FelcettoSheo SinghJun Wang , j
Agglutination－Assays
Agglutination AssaysREFERENCE: Lanyi, B., and T. Bergan. Methods in Microbiology, Vol 10: 93-168. BACTERIAL AGGLUTINATION: Bacterial agglutination is
Cellulase－assays
实验概要 Cellulase enzymes show activity during the ripening of some fruits, where their effects on cell walls results in softening of the frui
Carbohydrate－Assays
Carbohydrate AssaysREFERENCE: Wright and Rebers, Anal. Biochem. 49: 307-319, 1972.OBJECTIVE: To determine the relative amounts ofLPS carbohydrates pre
Cellulase－assays
Cellulase enzymes show activity during the ripening of some fruits, where their effects on cell walls results in softening of the fruit. In cases of p
夹心ELISA1
Enzyme-linked Immunosorbent Assays (ELISAs) combine the specificity of antibodies with the sensitivity of simple enzyme assays, by using antibodies or
The－Restriction－Enzyme－Database
REBASE The Restriction Enzyme Database The Restriction Enzyme data BASE A collection of information about restriction enzymes and related proteins. It
Restriction－Enzyme－Buffer
Restriction Enzyme Buffer Most enzymes can use REact buffers; however, some are made up separately. Use fresh Milli-Q water, siliconized or sterile gl
ATP1B2基因编码功能及结构描述
该基因编码的蛋白属于na+/k+和h+/k+atpaseβ链蛋白家族，属于na+/k+atpase亚家族。na+/k+-atpase是一种完整的膜蛋白，负责建立和维持na和k离子在质膜上的电化学梯度。这些梯度对于渗透调节、各种有机和无机分子的钠耦合传输以及神经和肌肉的电兴奋性是必不可少的。这种酶由两
ATP1B2－基因突变与药物因子介绍
该基因编码的蛋白属于na+/k+和h+/k+atpaseβ链蛋白家族，属于na+/k+atpase亚家族。na+/k+-atpase是一种完整的膜蛋白，负责建立和维持na和k离子在质膜上的电化学梯度。这些梯度对于渗透调节、各种有机和无机分子的钠耦合传输以及神经和肌肉的电兴奋性是必不可少的。这种酶由两
ATP1A1－基因突变与药物因子介绍
该基因编码的蛋白属于p型阳离子转运atp酶家族，属于na+/k+-atp酶亚家族。na+/k+-atpase是一种完整的膜蛋白，负责建立和维持na和k离子在质膜上的电化学梯度。这些梯度对于渗透调节、各种有机和无机分子的钠耦合传输以及神经和肌肉的电兴奋性是必不可少的。这种酶由两个亚单位组成，一个大的催
ATP1A1基因编码功能及结构描述
该基因编码的蛋白属于p型阳离子转运atp酶家族，属于na+/k+-atp酶亚家族。na+/k+-atpase是一种完整的膜蛋白，负责建立和维持na和k离子在质膜上的电化学梯度。这些梯度对于渗透调节、各种有机和无机分子的钠耦合传输以及神经和肌肉的电兴奋性是必不可少的。这种酶由两个亚单位组成，一个大的催
Fluorescent－Nucleoside－Triphosphates－for－SingleMolecule－Enzymology1
By: Christopher P. Toseland1 2 , Martin R. Webb1Affiliation(s): (1) MRC National Institute for Medical Research, London, UK(2) Institut für Zelluläre
Microtubule－Binding－Assays
MaterialsSiliconized ultracentrifuge microfuge tubesGTP-depleted microtubules6X SDS loading dye1X SDS loading dyeCoomassie Brilliant Blue R 250 (0.8%
Cytotoxicity－Assays－Protocol
Cytotoxicity Assays ProtocolCell-mediated cytotoxicity was determined by using a standard microcytotoxicity assay. Briefly, target cells were pelleted
Matrigel－invasion－assays
OverviewMatirgel is considered as basement membrane and generated from EHS sarcoma. Matrigel contains not only basement membrane components (collagens
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
Restriction－Enzyme－Digestion－of－DNA
Materials:10X restriction enzyme buffer (see manufacturer's recommendation)DNAsterile waterrestriction enzymephenol:chloroform (1:1)Add the follow
Enzyme－Kinetics－assay－of－the－WT
To assay 17 b-HSD activity in lysates, cells were harvested 48h after transfection using PBS Enzyme Free Cell Dissociation Solution ( specialty Media
Tubulin－Polymerization－with－GTP/GMPCPP/Taxol
I. Solutions & SuppliesII. Prepolymerization ClarificationIII. GTP PolymerizationIV. Taxol PolymerizationV. GMPCPP PolymerizationVI. Determining Conce
Solidstate－physics－offers－insights－into－dielectric－properties－of－...
Solid-state physics offers insights into dielectric properties of biomaterialsA team of Russian, Czech and German researchers gained a new perspecti
Activation－of－PKC－through－G－protein－coupled－receptor
G-protein coupled receptors (GPCRs) transduce a variety of signals from the extracellular environment across the plasma membrane. One of the common si
DNA－Fragmentation－Assays－for－Apoptosis
Protocol I: Triton X-100 Lysis BufferIn 96 flat-wells plate, incubate 4x10 6 target cells (40 wells of 105 per well) with desired concentration of eff