IsolationofMicrotubules(BovineBrain)
LEVEL IIMaterialsFreshly removed bovine brain 2Wire sieve (tea strainer)Microtubule buffer (MT buffer)0.1 M MES (2-(N-Morphilino)ethanesulfonic acid)1 mM EGTA (Ethylene Glycol-bis(-aminoethyl Ether) N,N,N',N'-tetraacetic acid)0.5 mM MgClAdjust pH to 6.48 M Glycerol in MT bufferVirtis homogenizer (or equivalent)Refrigerated centrifugeGTPBradford protein assayProcedure 3Remove the meninges and periphe......阅读全文
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
TEM－Visualization－of－Microtubules
LEVEL IIMaterialsCoated grid for TEM0.1 M ammonium acetate5% ethanol saturated uranyl acetateTransmission electron microscopeProcedureAt the conclusio
Viscosity－－Polymeriztion－of－Microtubules
LEVEL IIMaterialsTubulin (Brain extract from Exercise 9.4)GTPATPViscometerWaterbath or incubator at 37° CProcedureCompute the amount of GTP (M.W. 523)
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
细胞组分和细胞器——细胞器分离
Labeling Microtubules (Molecular Dynamics Inc. )Microtubules are involved in many aspects of cell motion including propulsion, mitosis, growth, and o
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
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
Ultraviolet－irradiation－impairs－epiboly－via－microtubules－in－Zebrafish
IntroductionZebrafish have transparentembryos that develop outside the mother. They develop rapidly, so that at 24 hours after fertilization, the embr
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/
Isolation－of－mouse－embryos
1. Sacrifice impregnated mouse.2. Dissect out the uterus of the mouse. Pulling up on the uterus with one set of forceps,use another to tear the mesome
mice－islet－isolation
1. Islets of Langerhans were isolated from 5- to 7-week-old nonobese diabetic (NOD) mice. 2. Which involves cannulation of the common bile duct and
Yeast－Nuclei－Isolation
This method gives yeast nuclei which look nearly purified microscopically. Nuclei isolated in this way do not give active transcription extracts when
Isolation－of－murine－splenocytes
OverviewIn order to study spleen cells (e.g. lymphocytes, granulocytes, other immune cells), it helps to make single-cell suspensions so that the cell
Isolation－of－liver－lymphocyte
Isolation of liver lymphocyte Several lymphocyte subpopulations reside in the normal adult human liver. Liver lymphocytes mainly include a large n
Isolation－of－papillary－cells
Isolation of renal papillary cells1. For isolation of papillary cells, kidneys were harvested and kept in HBSS containing 15 mM HEPES, penicillin/
Isolation－of－Zebrafish－embryos
Zebrafish will mate and deposit fertilized eggs on the bottom of the tank at 'dawn'. They can be accustomed to lay at any convenient time by k
Fungal－DNA－Isolation
Fungal DNA IsolationSaghai-Maroof MA, Soliman KM, Jorgensen RA, & Allard RW (1984) PNAS 81:8014-8018DNA successfully isolated from fungal species of C
Arabidopsis－gDNA－isolation
This is a simple and fast protocol for the extraction of genomic DNA from Arabidopsis thaliana that works fine in PCR for simple amplicons. We only us
Cosmid－DNA－Isolation
实验概要Isolation of high yields of highly pure cosmid DNA using PureLink™ HiPure Plasmid Purification Kits.实验原理The PureLink™ HiPure Plasmid Purification
Isolation－of－colonic－epithelium
The method we use is based on work of Dr. Hazel Cheng, at the University of Toronto and works for both colon and the small intestine.First we excise t
Chromosomal－DNA－Isolation
Chromosomal DNA IsolationMETHOD:Grow cells in 2-5 ml broth to late log phase.Pellet 1-2 ml cells in microfuge.Resuspend cells in 400 µl TES (50 mM Tri
Isolation－Of－PCR－Products
实验概要Rapid and efficient purification of PCR products from salts, primers, dNTPs, and other non-nucleic acid reagents.实验原理The ChargeSwitch® TechnologyT
DNA－isolation－extraction
CTAB TECHNIQUE / Method / Schedule / Protocol FOR DNA ISOLATION / DNA EXTRACTION FROM PLANT LEAF / LEAVES SAMPLES (see also DNA RNA double isolation
Isolation－of－papillary－cells
实验概要This protocols provides a general protocol for isolation of papillary cells.实验步骤Isolation of renal papillary cells1. For isolation of papillary c
RNA－isolation－for－Microarray
Description RNA extraction using TRI REAGENT. This method gives ample amout of RNA.Procedure It is 3 days procedure.Day 1:1. Harvest the cells and cen
Poly(A)+RNA－Isolation
Eukaryotic messenger RNA (mRNA) can be separated from the other RNA species in a total RNA preparation by affinity chromatography by virtue of the
RNA－Isolation－Protocol
Stabilize RNAStart with 15 ml E. coli Culture containing 7.5* 109 cells (OD600= 0.2 Dilute cells or scale up)Pipet 30 ml of RNAProtect Bacteria Reagen
Isolation－of－bone－marrow
(contributed by Chris Jackson (chris.jackson@bris.ac.uk))A protocol I have used to isolate rat bone marrow is: 1. Kill the rat and dissect out the fem