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......閱讀全文
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
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)
TEM－Visualization－of－Microtubules
LEVEL IIMaterialsCoated grid for TEM0.1 M ammonium acetate5% ethanol saturated uranyl acetateTransmission electron microscopeProcedureAt the conclusio
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
細胞組分和細胞器——細胞骨架
Fixation and Immunofluorescence of the Cytoskeleton?(Mitchison Lab)??Recycling Tubulin?(Mitchison Lab)??Labeling Tubulin and Quantifying Labeling Stoi
細胞組分和細胞器——細胞器分離
Labeling Microtubules?(Molecular Dynamics Inc.??)Microtubules are involved in many aspects of cell motion including propulsion, mitosis, growth, and o
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/
Nucleolar－Isolation－Protocol
We recommend that you first download and read this page as a?PDF file. Using that as your guide, you can then follow the protocol below and view a Qui
DNA－isolation－extraction
CTAB TECHNIQUE / Method / Schedule / Protocol FOR DNA ISOLATION / DNA EXTRACTION FROM PLANT LEAF / LEAVES SAMPLES (see also DNA RNA double isolation
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
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－liver－lymphocyte
Isolation of liver lymphocyte???Several lymphocyte subpopulations reside in the normal adult human liver. Liver lymphocytes mainly include a large n
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
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
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
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
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
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－papillary－cells
實驗概要This protocols provides a general protocol for isolation of papillary cells.實驗步驟Isolation of renal papillary cells1.?For ?isolation of papillary c
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
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
Isolation－Of－PCR－Products
實驗概要Rapid and efficient purification of PCR products from salts, primers, dNTPs, and other non-nucleic acid reagents.實驗原理The ChargeSwitch? TechnologyT
Cosmid－DNA－Isolation
實驗概要Isolation of high yields of highly pure cosmid DNA using PureLink? HiPure Plasmid Purification Kits.實驗原理The ?PureLink? HiPure Plasmid Purification
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
Isolation－of－colonic－epithelium
實驗概要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 e