FlowCellAssayswithMicrotubules:Motility／DynamicsinFluorescence
Flow cell assays are very useful for studying microtubule motility, microtubule dynamics, kinetochore-microtubule interactions and action of severing/depolymerizing factors on microtubules. Described here are some general procedures for flow cell assays.I. Solutions & SuppliesII. VE-DIC AssaysA. MotilityB. DynamicsIII. Fluorescence AssaysA. Oxygen ScavengingB. AssaysBack to protocolsI. Solutions & SuppliesBRB......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
Fluorescence－Procedures－forthe－Actin－andTubulin－Cytoskeleton－in－Fixed－Cells
Fluorescence Procedures for the Actin and Tubulin Cytoskeleton in Fixed CellsActin: Louise CramerTubulin: Arshad DesaiGeneral StrategyWe typically wor
Isolation－of－Microtubules－(Bovine－Brain)
LEVEL IIMaterialsFreshly removed bovine brain 2Wire sieve (tea strainer)Microtubule buffer (MT buffer)0.1 M MES (2-(N-Morphilino)ethanesulfonic acid)1
红细胞裂解的实验方法步骤
IntroductionPrior to using lymphoid tissue cell suspensions for flow cytometric analysis and/or for in vitro functional assays, it is recommended to r
线粒体荧光探针大全：TMRM，Mitotracker，JC1（3）
Mitochondrion-Selective Rhodamines and RosaminesRhodamine 123Rhodamine 123 (R302; FluoroPure Grade, R22420; Figure 12.22) is a cell-permeant, cationic
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
Fluorescence－Procedures－forthe－ActinandTubulin－Cytoskeleton－inFixed－Cells1
General StrategyWe typically work with tissue culture, primary mammalian cells, and cell extracts, but the protocols can be adapted to other systems,
Fluorescence－Mounting－Medium－(Antifade)
Materials Needed20ml glass scintillation vialSmall stir barFoilGlycerol1X PBSPipets* P-phenylenediamine ( EMD Chemicals Inc. Cat# PX0730)Carbonate-Bic
LIVE/DEAD®－Violet－Viability/Vitality－Kit
实验概要The LIVE/DEAD® Violet Viability/Vitality Kit provides a two-color fluorescence cell viability and vitality assay that is based on the simultane
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
DAPI－Counterstaining－Protocols
实验概要The blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; it appears to associate with AT clusters in the minor groove. Binding
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
Fixation－and－Embedding－of－Microtubules－for－Electron－Microscopy
(This procedure can also be used for virtually any material that must be pelleted prior to fixation and thin sectioning)Primary fix:2% glutaraldehyde
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
Negative－Stain－Electron－Microscopy－of－Microtubules
Negative staining is a rapid, qualitative method for analyzing microtubule structure at the EM level. Because negative staining involves deposition of
An－Integrative－Procedure－for－Apoptosis－Identification－and－Measurement2
TroubleshootingCritical Steps(1) Don’t trypsinize cells for too long when collecting them.(2) Rotation speed should be no more than 1500 rpm during ce
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－Cytometry－Analysis
PurposeFlow cytometry employs instrumentation that scans single cells flowing past excitation sources in a liquid medium. The technology can provide r
Fluorescence－In－Situ－Hybridization－using－TSA™
实验概要This protocol describes steps for fluorescent in situ hybridization (FISH) to Drosophila embryos using Tyramide Signal Amplification (TSA™), and
Apoptosis:－A－Laboratory－Manual－of－Experimental－Methods－Andrea－Cossarizza
THE CELL 1. Morphological aspects of apoptosis Walter Malorni, Stefano Fais & Carla Fiorentini 2. Cell cycle Miriam Capri & Daniela BarbieriTHE NUCLEU
An－Integrative－Procedure－for－Apoptosis－Identification－and－Measurement
IntroductionApoptosis is a normal physiological phenomenon put forward by Kerr [1]. It plays an important role in embryonic development, maintenance o
Application－Note:－Qdot®－Nanocrystal－Conjugates－in－Flow－Cytometry
实验概要Researchers today are trying to maximize the information that they get out of flow cytometry experiments by looking at more parameters in a sing
免疫荧光
Immunofluorescence Technique (Spector Lab)protocol for immunofluorescence on cells Immunofluorescence Protocol (Walter Steffen)Methanol fixationForma
DAPI－Nucleic－Acid－Stain
实验概要The blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; it appears to associate with AT clusters in the minor groove. Binding