MicrotubuleBindingAssays
MaterialsSiliconized ultracentrifuge microfuge tubesGTP-depleted microtubules6X SDS loading dye1X SDS loading dyeCoomassie Brilliant Blue R 250 (0.8% in 50% methanol + 10% acetic acid)Destaining solution (13% methanol + 3% acetic acid)Procedure1.Mix buffer and taxol in a siliconized ultracentrifuge microfuge tube, then add motor and GTP-depleted microtubules using a set concentration of microtubules (e.g., ......阅读全文
Microtubule－Binding－Assays
MaterialsSiliconized ultracentrifuge microfuge tubesGTP-depleted microtubules6X SDS loading dye1X SDS loading dyeCoomassie Brilliant Blue R 250 (0.8%
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
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
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
MinichromosomeMicrotubule－Binding－Assay－微染色体－微管结合实验1
Koshland Lab,Carnegie Institute http://www.ciwemb.edu/labs/koshland/Protocols/MICROTUBULE/mmb.htmlDetermine the OD600 and correlate the cell density f
MinichromosomeMicrotubule－Binding－Assay－微染色体－微管结合实验2
YWB per 10ml5mL 2M Sorbitol (if NZ arrested, add 40uL 1.5mg/mL0.336mL 1M K2HPO4 N2 to 4mL YWB)0.064mL 1M KH2PO44.6mL dH2OYWB, glycerol, PMSF5mL 2M Sor
Agglutination－Assays
Agglutination AssaysREFERENCE: Lanyi, B., and T. Bergan. Methods in Microbiology, Vol 10: 93-168. BACTERIAL AGGLUTINATION: Bacterial agglutination is
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
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
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
Glycolipid－Binding－Assay
Glycolipid Binding AssaySource: Contributed by Pingsunjim, Paller’s LabAbstract: This protocol can be used for the detection of glycolipids binding to
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
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
Nucleotide－Binding/Hydrolysis－Assay
MaterialsNucleotide mixMotor (50 - 100 µM; purity > 95%)0.5 M Tris-OAc, pH 7.510 mM EGTA10 mM MgCl2DDWSephadex G-50 Medium column (0.8 cm in x 20 cm)C
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
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/
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
IASYSbinding－cuvette－and－getting－KD
Immobilization of ligands on cuvette surfaces and measure the interactions of ligand which is immobilized on the cuvette and the ligate which is a
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
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
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
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
实验室自动化与筛选协会2013亚洲会展短期培训课程简介
短期培训课程I ：生物制剂研发中的高通量筛选 Junma Zhou，博士，上海药明康德新药开发有限公司 刘洁颖博士，上海药明康德新药开发有限公司 In Scope of Short Course - Overview of high-throughput techniques in