Microtubule(MT)／OrganelleMotilityAssays
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 regeneration system. Dilute the axonemes with PM Buffer to the proper dilution (determined after preparation, see support protocol 2 below) and prepare 6x membranes (determined after preparation, see support protocol 4 below) by diluting organelles with PM buffer contain......阅读全文
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
Microtubule－Binding－Assays
MaterialsSiliconized ultracentrifuge microfuge tubesGTP-depleted microtubules6X SDS loading dye1X SDS loading dyeCoomassie Brilliant Blue R 250 (0.8%
Flow－Cell－Assays－with－Microtubules:－Motility/Dynamics－in－Fluorescence
Flow cell assays are very useful for studying microtubule motility, microtubule dynamics, kinetochore-microtubule interactions and action of severing/
Motility－Assay
DescriptionVarious phenotypic characteristics are requiredfor a cancer cell to successfully complete the metastaticcascade. Among these, acquisition o
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
Microtubule－Spindowns－for－Visual－Analysis
Microtubule spindowns for visual analysis can be performed on single microtubules or microtubules nucleated from axonemes/centrosomes. Although live D
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 3.0x
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
mCalpain－and－friends－in－Cell－motility
The mammalian calpain gene family currently contains 13 distinct large subunit products most of which complex with one of two smaller 30kDa subunits.
Matrigel－invasion－assays
OverviewMatirgel is considered as basement membrane and generated from EHS sarcoma. Matrigel contains not only basement membrane components (collagens
Cytotoxicity－Assays－Protocol
Cytotoxicity Assays ProtocolCell-mediated cytotoxicity was determined by using a standard microcytotoxicity assay. Briefly, target cells were pelleted
Rho－cell－motility－signaling－pathway
RhoA is a small G-protein in the Rho family that regulates cell morphology via actin cytoskeleton reorganization in response to extracellular signals.
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
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
PLAQUE－ASSAYS－FOR－ADENOVIRUS－TITRATION
-Set up 60 mm dishes of P11 cells to be 100 confluent at time of infection. -Remove medium from dishes, add 0.2 to 0.5 ml virus and adsorb for 30 – 60
cAMP分析－cAMP－Assays
cAMP AssaysGouzel Karimova and Daniel LadantUnite Postulante de Biochimie des Interactions Macromoleculaires, Departement de Biologie Structurale et C
SAPK/Jun－kinase－assays
Preparation of cell lysate:1. For cells in suspension, grow in DMEM with 10% FCS and then the day before you do the experiment spin the cells down (1
Coimmunoprecipitation－assays
co-IP assays can be performed between endogenous proteins or transiently or stably expressed exogenous - usually tagged - proteins. The advantage to u
Rac－1－cell－motility－signaling－pathway
Rac-1 is a small G-protein in the Rho family that regulates cell motility in response to extracellular signals. Several changes in cytoskeletal struct
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
Fragment－Complementation－and－Coimmunoprecipitation－Assays－for－...
Plant disease resistance (R) proteins confer protection against specific pathogens or pathogen isolates. R proteins function by recognizing pathog
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
Preparation of Luciferin forIn Vitro Bioluminescent AssaysMaterials• D-Luciferin Firefly, potassium salt, 1.0 g /vial(Caliper Life Sciences Part Numbe
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
实验概要Reagent for immunoassay, ligand binding assay and ligand receptor assay in which luciferin is covalently bonded to a molecule having biological
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
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
实验概要Reagent for immunoassay, ligand binding assay and ligand receptor assay in which luciferin is covalently bonded to a molecule having biological
Tubulin－Polymerization－with－GTP/GMPCPP/Taxol
I. Solutions & SuppliesII. Prepolymerization ClarificationIII. GTP PolymerizationIV. Taxol PolymerizationV. GMPCPP PolymerizationVI. Determining Conce
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