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
BackgroundThis method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to ente
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 which are expressed and then degraded in a concerted f
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
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
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
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－ProtocolDAPI
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.
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
Regulation－of－cell－cycle－progression－by－Plk3
The focus of this pathway is to illustrate the role of Polo-like Kinase 3 (Plk3 also known as Prk and Fnk) as part of the regulatory cascade leading t
Cell－Cycle:－G2/M－Checkpoint
The G2/M DNA damage checkpoint prevents the cell from entering mitosis (M phase) if the genome is damaged. The Cdc2-cyclin B kinase is pivotal in regu
Cell－Cycle：细胞永生化研究进展
每一天，你身体内的一些细胞会停止分裂，这是一件好事。而有些细胞会无限增殖，这是大多数恶性肿瘤发展的一个重要早期步骤。 尽管细胞无限增殖在癌症中有着非常的重要性，但研究人员一直对于细胞永生化背后的分子机制知之甚少。这是因为科学家缺乏好方法研究永生化人类细胞。在发表在Cell Cycle 杂志上的
Cell－Cycle:－G1/S－Check－Point
The G1/S cell cycle checkpoint controls the passage of eukaryotic cells from the first 'gap' phase (G1) into the DNA synthesis phase (S). Two
Regulation－of－p27－Phosphorylation－during－Cell－Cycle－Progression
p27/Kip1 regulates the cell cycle by inhibiting the checkpoint kinase cdk2/cyclin E and blocking cell cycle progression through the G1-S transition. C
Cell－Cycle－－J－Neurosci：脊髓损伤引发大脑损害
很多脊髓损伤的研究都集中于揭示脊髓损伤所引发的效应，然而往往在研究中科学家们很容易忽视脊髓损伤对大脑所引发的效应。近日，来自马里兰大学的研究人员通过研究首次揭示，脊髓损伤（SCI）可以促进广泛和持续的大脑炎症，进而引发神经细胞的逐渐缺失以及认知问题和抑郁症等。 相关研究发表于国际杂志Journ
Sonic－Hedgehog－(SHH)－Receptor－Ptc1－Regulates－cell－cycle
Sonic Hedgehog (Shh) is a secreted protein identified genetically as an important developmental factor. Shh provides a morphogenic signal in the devel
Estrogenresponsive－protein－Efp－controls－cell－cycle－andbreast－tumors－growth
The estrogen-inducible RING finger protein Efp stimulates proliferation of breast cancer cells. 14-3-3 is p53-inducible protein that sequesters mitoti
细胞周期试剂盒Cell－cycle－staining－Kit中文操作步骤
试剂盒特点：1、独创的活细胞一步法检测，收集——洗涤——染色，简单快速 2、兼容固定细胞检测，改进的固定方法，更少细胞粘连，更小CV值 3、兼容流式和荧光显微镜分析，提供详细protocol固定细胞处理：1、收集细胞悬液，细胞数为2 × 10^5 ~ 1 × 10^6。离心沉淀细胞，去上清，
Cell－Cycle：乳腺癌细胞耐药性的分子机理
近日，来自Norris Cotton综合癌症研究中心的研究人员通过研究开发了一种治疗ERBB2（人类表皮生长因子受体II）阳性乳腺癌的新型疗法，该类乳腺癌通常会对疗法产生强烈的耐药性，相关研究刊登于国际杂志Cell Cycle上，该研究为揭示新型的癌症耐药机制提供了一定的线索。 研究者Kuro
The－Citric－Acid－Cycle
The Krebs cycle, also called the citric acid cycle, is a fundamental metabolic pathway involving eight enzymes essential for energy production through
Method:－Reactivating－Cell－Lines－and－Cell－Growth－for－DNA－Preparation
Purpose:Cell lines are reactivated and grown to a count of 1 x 108 cells. The cells are pelleted and stored frozen at -80 degrees C prior to DNA extra
Cosmid－Cloning:－Cell－preparation,－DNA－packaging,－and－Cell－Transfection
Cosmid Cloning: Cell preparation, DNA packaging, and Cell TransfectionProtocol taken from Stratagene's Gigapack packaging extracts instruction man
RB－Tumor－Suppressor/Checkpoint－Signaling－in－response－to－DNA－damage
Cell cycle checkpoint controls at the G1 to S transition and the G2 to M transition prevent the cell cycle from progressing when DNA is damaged. The A
The－SARScoronavirus－Life－Cycle
Severe Acute Respiratory Syndrome (SARS) has affected thousands of individuals, causing fever, and pneumonia with a mortality rate estimated at 10-18%
CDK－Regulation－of－DNA－Replication
Initiation of DNA replication in eukaryotes is a highly conserved, multi-step process (replication licensing) designed to restrict initiation events t
Cell：DNA损伤应答的新通路
加州大学的科学家们在研究DNA损伤对高尔基体的影响时，发现了DNA损伤激活的新通路，这一通路影响着机体中细胞对化疗的应答。 包括化疗和放疗在内的标准癌症治疗策略，通过诱导细胞出现DNA损伤起作用。DNA损伤启动的信号通路会导致细胞死亡，人们正是在这一机制的基础上消灭癌细胞。更好地理解这些细
《Cell》探讨“垃圾”DNA的新功能
美国国立卫生研究院下属国家关节肌肉骨骼及皮肤病研究所(NIAMS)的科学家们在新研究中证实，过去被认为是“垃圾”的DNA在免疫系统反应中发挥了关键性的作用。研究结果发表在近期的《细胞》(Cell)杂志上，有可能促成发现治疗免疫相关疾病的新靶点。 人类基因组中有32亿DNA碱基对，但只有2%
Cell：超越DNA的遗传与编程
来自犹他大学Huntsman癌症研究所(HCI)的研究人员在新研究中发现，父源基因在受精之时已预编程至胚胎所需状态，而母源基因则处于另一种状态，还必须进行重编程才能与之相匹配。这一研究发现对于发育生物学和癌症生物学均具有极其重要的意义。研究论文发表在5月9日的《细胞》(Cell)杂志上。
Cell：DNA损伤揭示抗癌新疗法
大自然中每一个有机体都会不惜代价保护自身的DNA，但细胞如何精确区分自身DNA的损伤还是入侵病毒外源DNA的损伤依然是个谜底，近日刊登在国际杂志Cell上的一篇研究论文中，来自索尔克研究所的研究人员通过研究揭示了细胞反应系统精确区分上述两种威胁的机制，相关研究或可帮助开发新型的癌症选择性病毒疗法