DNACellCycle
Solutions70% ethanolribonuclease (100 µg/ml DNase free, Sigma)propidium iodide ( 50 µg/ml in PBS)ProcedureHarvest cells. Spin at 1200 rpm for 5 minutes.Discard supernatant; add 1 ml ' fridge cold' 70% ethanol to 1x106 cells while vortexing the cell pellet gently.Fix for at least 30 min. at 4° C.Spin at 2000 rpm for 5 min. Once in 70% ethanol, samples may be kept for up to 2 weeks.Discard supernatant and resus......阅读全文
DNA Cell Cycle
Solutions70% ethanolribonuclease (100 µg/ml DNase free, Sigma)propidium iodide ( 50 µg/ml in PBS)ProcedureHarvest cells. Spin at 1200 rpm for 5 minute
FIXATION and DNA Staining for Cell Cycle Analysis
Background This method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to e
CELL CYCLE ANALYSIS
PROPIDIUM IODIDE: The most commonly used dye for DNA content/cell cycle analysis is PROPIDIUM IODIDE (PI). It can be used to stain whole cells or isol
Cyclins and Cell Cycle Regulation
The cell cycle is regulated by the interplay of many molecules. Key among these are the cyclins whi
Flow Cytometric Analysis of Cell Cycle
Fixation1) Collect 2 X 106 cells.2) Pellet cells by spinning at 1,000 rpm, 4°C for 5 minutes.3) Resuspend cell pellet in 1 ml of cold PBS.4) Fix cells
Yeast Cell Cycle by Flow Cytometry
ReagentsCold absolute ethanol.0.5 M Na citrate stock (filtered), 50mM diluted stock.10 mg/ml RNase A (Boil 10 mins, cool, filter and store at -20°C).4
PTEN dependent cell cycle arrest and apoptosis
PTEN is a tumor suppressor gene. Recombinant PTEN is capable of dephosphorylating phosphatidylinositol 3,4,5-triphosphate[PI(3,4,5)P3], the product of
Cell Cycle Staining Protocol-DAPI
1. Harvest cells- wash 2X in PBS to get rid of serum proteins. 1200rpm, 5 min2. Resuspend pellet (up to 3x106 cells) in 1.2 ml PBS (Ca and Mg free).3.
Cell cycle analysis of Escherichia coli cells
Cell cycle analysis of Escherichia coli cellsC period = the time for a round of chromosome replicationD period = the time between the end of a round o
BTG family proteins and cell cycle regulation
BTG2 is found to be one of the immediate early genes up-regulated by neural growth factor (NGF) and epidermal growth factor (EGF). Its transcriptional