细胞功能测定

DNA Fragmentation Assays for Apoptosis ProtocolProtocol I: Triton X-100 Lysis Buffer In 96 flat-wells plate incubate 4x10 6 target cells (40 wells of 105 per well) with desired concentration of effect ...

Staining Methods for cell death Z. Xia 10/2/95The simplest way: trypan blue. Dead cells stain blue Non-fixed cells: FDA(fluorescein diacetate)-green alive cells; P.I. (propidium iodide)-red dead cells ...

In Situ Cell Death (Apoptosis) Detection by TUNEL labelingby Boehringer Mannheim (Catalog No. 1684809) modified by Josiah N. Wilcox andJosé C. Rodriguez - Emory University - April 1996Protocol for Fro ...

Detection of Apoptosis in Paraffin-embedded tissues by ISEL (In Situ End Labeling)The ISEL technique identifies morphologically apparent karyorrhetic cells and also labels cells in which the DNA frag ...

Paraformaldehyde Fixation of CellsBackgroundThis fixation method is good for cells labelled by fluorochrome-conjugated antibodies to membrane antigens. It will stabilize the light scatter and labellin ...

Basic Method for Direct Immunofluorescence Labeling BackgroundThis is the method for direct immunofluorescence labeling; that is the antibodies have the fluorescent dye attached. Direct labeling is si ...

BRB80 (1X): 80 mM PIPES 1 mM MgCl2 1 mM EGTA pH 6.8 with KOH (generally made as a 5X stock and stored at 4¡C) 100 mM GTP 100 mM GMPCPP Taxol : 10 mM stock ...

Viscosity & Polymeriztion of MicrotubulesLEVEL II Materials Tubulin (Brain extract from Exercise 9.4)GTPATPViscometerWaterbath or incubator at 37° C Procedure Compute the amount of GTP (M.W. 523) need ...

Flow cytometric determination of leukocyte surface antigens in whole bloodThis document assumes that you have a mouse IgG monoclonal antibody directed to the antigen you wish to quantitate and that yo ...

FIXATION and DNA Staining for Cell Cycle Analysis BackgroundThis method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane which allows the dye (Propidium Iodide) to enter ...

MT Spindowns from Extracts Arshad DesaiNotes:The key variable in MT spindown experiments is ATP. Under high ATP conditionsconventional MAPs are selectively co-sedimented with microtubules. In theabsen ...

Fluorescein labeling of proteinsThis method uses carboxyfluorescein succinimidyl ester (CFSE) rather than fluorescein isothiocyanate resulting in more reliable labeling. Succinimidyl esters are excell ...

Microtubule Spindowns for Visual Analysis Microtubule spindowns for visual analysis can be performed on single microtubules or microtubules nucleated from axonemes/centrosomes. Although live DIC analy ...

Flow Cytometry Assayto Measure Internalization of GPCR's contributed by Patricia TsaoInternalization of receptor is measured as the reduction of receptors on the plasma membrane surface. Surface recep ...

Flow Cytometry: Immunofluorescence Staining of Activated and Resting Human PlateletsPROCEDURE for preparation of activated platelets1. Centrifuge tube of freshly drawn HEP or EDTA blood @ 600 rpm (75x ...

I. Useful Values 1 mg/ml tubulin = 10 µM (assuming MW of ab-tubulin heterodimer is 100000; in reality it is ~110000 but almost all tubulin labs use this convenient conversion relationship) 1 µm of a m ...

Although many protocols for tubulin preparation are available the procedure described below is the simplest and highest yielding preparation I have done. The protocol calls for 3 pig brains and shoul ...

Passaging cellsPour out media from flasks. Wash with Hanks. 5 ml per flask. Tilt around then dump. Add 4 ml of Trypsin / EDTA to each flask. Tip then bang. Add 20% FBS NCTC media and tilt. 4 ml per fl ...

Cell counting with an hemacytometer.Accuracy of manual counts with an hemacytometer depend on: accurate mixing of the sample (no bubbles!) number of chambers counted number of cells counted (practica ...

Immunodetection of cyclin D1 and D2/D3 using 488/630 nm dual laser flow cytometry William Telford Hospital for Special Surgery This protocol is for use with the D cyclins and employs 488 nm argon las ...