细胞功能测定

Stewart Laboratory Standard Operating Procedure BrdU Incorporation ProtocolDay 1Label cells with media containing100 μM BrdU (6 μL/mL of 5 mg/mL stock): Diploid Fibroblasts 4-6 hours ES ...

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 ...

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 ...

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 ...

Flow Cytometric Analysis of Cell CycleContributor: Suprya JayadevDate: July 13 1994Fixation1) Collect 2 X 106 cells.2) Pellet cells by spinning at 1000 rpm 4°C for 5 minutes.3) Resuspend cell pellet i ...

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 ...

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 ...

Authentication TechniquesWhatever the scope of work to be carried out it is important to know that the work is being conducted using the correct reagents. This is no less important for cell cultures s ...

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 ...

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

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 ...

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 ...