细胞功能测定

Immunofluorescence Staining of Human Cells by Lysed Whole Blood MethodAdd 100 m l of well-mixed anticoagulated whole blood to the bottom of a labeled tube. Add the appropriate primary antibody to each ...

General Annexin V Staining ProcedureSolutions 1. 10X Binding Buffer (Cat. No. 66121A) : 0.1 M HEPES pH 7.4; 1.4 M NaCl; 25 mM CaCl 2 . Dilute to 1X prior to use. 2. Propidium Iodide (PI). Prepare a ...

Fluorescence Procedures for the Actin and Tubulin Cytoskeleton in Fixed CellsActin: Louise CramerTubulin: Arshad DesaiGeneral StrategyWe typically work with tissue culture primary mammalian cells and ...

Flow Cell Assays with Microtubules: Motility/Dynamics in Fluorescence and VE-DIC Flow cell assays are very useful for studying microtubule motility microtubule dynamics kinetochore-microtubule interac ...

Labeling Tubulin and Quantifying Labeling StoichiometryThis is a general procedure for coupling moieties with reactive succinimidyl esters to tubulin. We have used it successfully to derivatize tubuli ...

Recycling TubulinWe "recycle" tubulin fractions stored at -80¡C after the PC column and store the recycled tubulin in small aliquots for day-to-day use. We generally store recycled tubulin in Injectio ...

SDS Gel Electrophoresis of Tubulin\MAPsLEVEL II Figure 9.3 SDS-PAGE of tubulin and associated proteins Materials Stock Acrylamide: (30%T:0.8%C)30% by weight of acrylamide0.8% by weight of NN'-bis-meth ...

TEM Visualization of MicrotubulesLEVEL II Materials Coated grid for TEM0.1 M ammonium acetate5% ethanol saturated uranyl acetateTransmission electron microscope Procedure At the conclusion of a 37° C ...

Cell Disruption and Subcellular FractionationContributor: Suprya JayadevDate: July 31 1991Nitrogen Bomb1) Harvest cells and wash 2 times with PBS.2) Resuspend final pellet in relaxation buffer. (20 ml ...

CHO Centrosome Prep:Arshad Desai4/94Cells:We grow our CHOs with MEM (without nucleosides) + 10% Bovine Calf Serum and penn/strep/glutamine. For a prep it is best to grow twenty large plates (150 mM) a ...

Lysosome Isolation in Isotonic SucroseLEVEL I MATERIALS Rat liverPhysiological saline (0.85% w/v NaCl)0.25 M sucrose in 10 mM Tris-HCL pH 7.4Brendler teflon homogenizerRefrigerated preparative centrif ...

Organelle DNA Library Construction(version MAY-1998)I. NEBULIZATION of DNA 1. 0.5 - 5 ug DNA in TE (10mM/1mM) 25% glycerol final volume 500 ul 2. nebulize for 90-100 sec at 5-10 psi (for a med ...

Preparation of Mitochondria from Rat Liver Rat liver is an ideal source for functional intact mitochondria for a number of reasons. We use Sprague-Dawley albino male rats for our studies. Female rats ...

Recommended Experiments with Isolated Mitochondria Each oxygraph setup will consist of two stations each with dissolved oxygen chamber and chart recorder connected to a two-channel oxygen monitor. Ple ...

Mitochondria Structure and Function - Theoretical DetailsIn an attempt to be concise and understandable introductory level courses and textbooks frequently present concepts that are technically correc ...

Cytotoxicity Assays ProtocolCell-mediated cytotoxicity was determined by using a standard microcytotoxicity assay. Briefly target cells were pelleted and resuspended in 100 mCi of Na51Cr (DuPont NEN B ...

Phycoerythrin conjugation protocolDavid's method modified from references (2) and (3). I used this method to conjugate a mouse IgG2a monoclonal antibody (IB4). It works well. ReagentsAbAntibody about ...

Adrenal chromaffin granule (chromaffin vesicle) preparationIntroduction. This prep is adapted from the classic paper of Smith and Winkler (Smith AD; Winkler H. A simple method for the isolation of ad ...

Pheromone Halo AssayREFERENCE: Hoffman G. Garrison T. R. and Dohlman H. G. Analysis of RGS proteins in Saccharomyces cerevisiae Methods Enzymol.344:617-631 2002. -Use sterile technique and sterile s ...

Overview This assay uses a simple apparatus to study chemotaxis of leukocytes or other migratory cells. The apparatus consists of two multi-well chambers separated by a filter containing pores of uni ...