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......閱讀全文
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
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
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
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.
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:－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
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:－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
Cell－Cycle：細胞永生化研究進展
每一天，你身體內的一些細胞會停止分裂，這是一件好事。而有些細胞會無限增殖，這是大多數惡性腫瘤發展的一個重要早期步驟。 盡管細胞無限增殖在癌癥中有著非常的重要性，但研究人員一直對于細胞永生化背后的分子機制知之甚少。這是因為科學家缺乏好方法研究永生化人類細胞。在發表在Cell Cycle 雜志上的
Cell－Cycle－－J－Neurosci：脊髓損傷引發大腦損害
很多脊髓損傷的研究都集中于揭示脊髓損傷所引發的效應，然而往往在研究中科學家們很容易忽視脊髓損傷對大腦所引發的效應。近日，來自馬里蘭大學的研究人員通過研究首次揭示，脊髓損傷（SCI）可以促進廣泛和持續的大腦炎癥，進而引發神經細胞的逐漸缺失以及認知問題和抑郁癥等。 相關研究發表于國際雜志Journ
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
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
Cell－Cycle：乳腺癌細胞耐藥性的分子機理
近日，來自Norris Cotton綜合癌癥研究中心的研究人員通過研究開發了一種治療ERBB2（人類表皮生長因子受體II）陽性乳腺癌的新型療法，該類乳腺癌通常會對療法產生強烈的耐藥性，相關研究刊登于國際雜志Cell Cycle上，該研究為揭示新型的癌癥耐藥機制提供了一定的線索。 研究者Kuro
細胞周期試劑盒Cell－cycle－staining－Kit中文操作步驟
試劑盒特點：1、獨創的活細胞一步法檢測，收集——洗滌——染色，簡單快速 ? 2、兼容固定細胞檢測，改進的固定方法，更少細胞粘連，更小CV值 ? 3、兼容流式和熒光顯微鏡分析，提供詳細protocol固定細胞處理：1、收集細胞懸液，細胞數為2 × 10^5 ~ 1 × 10^6。離心沉淀細胞，去上清，
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
光合碳循環－（photosynthetic－carbon－cycle）
光合作用中碳同化（二氧化碳轉化為糖或其磷酸酯）的基本途徑。又稱卡爾文循環、還原戊糖磷酸循環、還原戊糖磷酸途徑。在綠色植物、藍藻和多種光合細菌中普遍存在。其他碳同化途徑如 C4 途徑和 CAM途徑（見景天科酸代謝）所固定的 CO2 ，最終仍須通過光合碳循環才能被還原成糖。因此它是地球上絕大部分有機物
The－reactions－that－feed－amino－groups－into－the－urea－cycle
Excess amino acids in the body can be used as a source of energy, with their carbon skeleton converted to metabolic intermediates such as acetyl-CoA o
Cell：DNA損傷應答的新通路
加州大學的科學家們在研究DNA損傷對高爾基體的影響時，發現了DNA損傷激活的新通路，這一通路影響著機體中細胞對化療的應答。 包括化療和放療在內的標準癌癥治療策略，通過誘導細胞出現DNA損傷起作用。DNA損傷啟動的信號通路會導致細胞死亡，人們正是在這一機制的基礎上消滅癌細胞。更好地理解這些細
《Cell》探討“垃圾”DNA的新功能
美國國立衛生研究院下屬國家關節肌肉骨骼及皮膚病研究所(NIAMS)的科學家們在新研究中證實，過去被認為是“垃圾”的DNA在免疫系統反應中發揮了關鍵性的作用。研究結果發表在近期的《細胞》(Cell)雜志上，有可能促成發現治療免疫相關疾病的新靶點。 人類基因組中有32億DNA堿基對，但只有2%