MicrotubuleSpindownsforVisualAnalysis
Microtubule spindowns for visual analysis can be performed on single microtubules or microtubules nucleated from axonemes/centrosomes. Although live DIC analysis has largely superseded the use of visual fixed time point assays for analyzing microtubule dynamics, spindowns can still be very useful. Procedures for fixing and pelleting microtubules onto coverslips are described here.I. Solutions & SuppliesII. Spindo......阅读全文
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
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
Visual－Signal－Transduction
The signal transduction cascade responsible for sensing light in vertebrates is one of the best studied signal transduction processes, and is initiate
MT－Spindowns－from－Extracts
MT Spindowns from ExtractsArshad DesaiNotes:The key variable in MT spindown experiments is ATP. Under high ATP conditions,conventional MAPs are select
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
Glycosphingolipid－analysis
1) Incubate cells with 1 µCi/ml of 3H-galactose for 72 hours.---> If treatment is for an extended period of time: treat in serum free media containing
Lipid－analysis
Thin layer chromatography is based on the separation of a mixture of compounds as it migrates with the help of a suitable solvent through a thin layer
Analysis－of－Heme－and－Hemoproteins
Heme is perhaps the most ubiquitous cofactor found in nature and the most functionally diverse. Hemoproteins are involved in cell respiration (cyt
Flow－Cytometry－Analysis
PurposeFlow cytometry employs instrumentation that scans single cells flowing past excitation sources in a liquid medium. The technology can provide r
Lineage－Analysis－of－Blood
Materials:Capillary tubes1.5 mL Eppendorf microfuge tubes15 mL conical centrifuge tubes96-well V-bottom plates (Corning Costar 3894, from Fisher)Flow
Biosynthesis－and－Analysis－of－Bilins
The term bilin is a collective one to describe a broad group of open chain tetrapyrroles and derives from the name “bile pigments” as the first of
Analysis－of－Oligosaccharide－Ligands
Analysis of Oligosaccharide Ligands by High Performance Liquid Affinity ChromatographyAnalysis of Oligosaccharide Ligands by High Performance Liquid A
CELL－CYCLE－ANALYSIS
PROPIDIUM IODIDE: The most commonly used dye for DNA content/cell cycle analysis is PROPIDIUM IODIDE (PI). It can be used to stain whole cells or isol
Analysis－of－murine－BM
Histologic analysis of murine BM is a necessary complement to flow cytometric or in vitro analysis. Techniques to do this are well established in huma
Analysis－and－Reconstitution－of－Phycobiliproteins:－Methods－for－the－...
Analysis and Reconstitution of Phycobiliproteins: Methods for the Characterization of Bilin Attachment ReactionsPhycobiliproteins are a homologous fam
Molecular－Analysis－and－Results－－DNA
Theory of CGHComparative genomic hybridization (CGH) is a fairly new molecular cytogenetic technique that allows detection of DNA sequence copy number
FACS－Analysis－of－ES－Cells
Isolate cells and dissociate to single cell suspension (can use Gibco Cell Dissociation Buffer, Accutase or Trypsin)Wash with 10% FBS/DMEM:F12For surf
Protocols－for－LCM－preparation－and－analysis
Protocols for LCM preparation and analysis I. Preparation, LCM and RNA/DNA extraction of Frozen Tissue SectionsA. EmbeddingB. CuttingC. StainingII. Pr
Genomic－Southern－Blot－Analysis
This chapter describes a detailed protocol for genomic Southern blot analysis which can be used to detect transgene or endogenous gene sequences i
PAM:－Prediction－Analysis－for－Microarrays
PAM: Prediction Analysis for MicroarraysClass Prediction and Survival Analysis for Genomic Expression Data Mining Features:Performs sample classificat
Simultaneous－analysis－of－DNA－content
Simultaneous analysis of DNA content and surface immunophenotype using gentle ethanol fixation techniques. William Telford. Louis E. King and Pamela
SOLID－TUMOR－CULTURES－FOR－CHROMOSOME－ANALYSIS
I. Purpose:A. Samples of solid tumors or lymph nodes may be sent from patients with cancer. These samples should be processed directly and also set up
TISSUE－FIBROBLAST－CULTURES－FOR－CHROMOSOME－ANALYSIS
I. Purpose:A: Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. suspected
Flow－Cytometric－Analysis－of－Cell－Cycle
Fixation1) Collect 2 X 106 cells.2) Pellet cells by spinning at 1,000 rpm, 4°C for 5 minutes.3) Resuspend cell pellet in 1 ml of cold PBS.4) Fix cells
TISSUE－FIBROBLAST－CULTURES－FOR－CHROMOSOME－ANALYSIS
实验概要 Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. suspect
Lipoprotein－Analysis－Week－2:－Electrophoresis
Lipoprotein Analysis Week 2: Electrophoresis IntroductionSDS polyacrylamide gel electrophoresis (SDS PAGE) will be used to assess the purification pr
Reverse－Transfection－for－Gene－Function－Analysis
This guide describes a microarray-based system for the functional analysis in mammalian cells of many genes in parallel. Mammalian cells are cultured