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2020 - 05 - 08
最近,《自然—通讯》刊发了一篇针对类风湿关节炎(RA)致病和标志分子的研究。该文章来自北京大学人民医院风湿免疫研究所栗占国课题组,作者发现,RA患者的sSR-A水平显着升高,与该疾病的临床症状和免疫状况指标直接相关。结果表明sSR-A是RA诊断的新型生物标志物,尤其可用于血清阴性及早期RA的诊断,而靶向sSR-A可能是一种新的治疗策略。《中国科学报》了解到,RA是一种常见的自身免疫病,具有高致残性。全球有3000多万患者,该病以破坏性关节炎及自身免疫异常为主要特征。目前,RA的血清诊断主要依据类风湿因子和抗CCP的检测。但是,仍有大量患者在发病初期,甚至患病多年而缺乏生物标志物,给临床诊断和治疗带来困难,导致延误治疗而出现残疾。早期诊断对于改善RA的结局至关重要。但是,目前的诊断工具对早期患者敏感性有限,致使大量的患者因延误诊断,不能得以正确治疗,出现残疾。鉴于此,该研究团队联合浙江大学、南方医科大学和包头医学院的研究者展开了一项大规模的多中心研究,涉及3262名参与者的训练队列和验证队列,最终发现sSR-A是RA的致病分子,可促进炎性T细胞和致炎因子产生。并且,该课题组利用基因敲除的sSR-A-/-模型研究证明,sSR-A可以促进关节炎的发病以及病情的加重,靶向抑制sSR-A可以缓解关节炎的症状。研究者进一步证实,sSR-A蛋白可以促进胶原诱导性关节炎发病,加速病情进展,加重疾病...
2
2019 - 08 - 01
第二看台老年性黄斑变性(AMD)、视网膜色素变性(RP)等视网膜变性疾病是目前尚无有效治疗手段的凶险致盲性眼病,也是公认的 “视力杀手”。由中南大学爱尔眼科学院院长唐仕波教授、爱尔眼科研究所副所长陈建苏教授带领团队,利用诱导多能干细胞(iPS)条件培养基和飞秒角膜透镜,联合构建高活性视网膜色素上皮细胞(RPE)的研究,不但有利于对视网膜色素上皮细胞生理特性、药物筛选等研究,还有利于构建组织工程视网膜层及其移植。这将帮助科学家开发出应用组织工程 RPE 细胞片治疗视网膜变性疾病和光感受器病变的新方法。培养高活性的视网膜细胞与年龄相关的老年性黄斑变性是发达国家 50 岁以上人群视力损害和致盲的首要病因,多达 90% 的病人无法治愈。数据显示,发达国家年龄 50 岁以上黄斑变性的发病率达 11%,目前我国的发病率接近发达国家,患病人数超过 3000 万人,并以每年 30 万人次的速度增加,成为我国现阶段 50 岁以上年龄群体发病率最高的三大眼科疾病之一,而且女性呈现出比男性更高的危险性。研究发现,未来最有希望治疗老年性黄斑变性的方式是视网膜色素上皮细胞疗法、视网膜黄斑部分的移植。那么,要采用 RPE 疗法,就需要先培养健康的视网膜细胞。陈建苏告诉科技日报记者,取出死亡时间在 3 小时内捐献者的带有部分脉络膜的视网膜组织,在解剖显微镜下可将带有部分脉络膜的 RPE 层从捐赠组织中分离出来,...
3
2019 - 07 - 26
科技日报北京 7 月 30 日电 (记者付丽丽)“建立严格的法律责任制度,对违反《疫苗管理法》规定构成犯罪的,依法从重追究刑事责任,货款不足 50 万元的按 50 万元计算。”30 日,在国家药监局主办的 “《疫苗管理法》颁布法规解读宣贯研讨会上”,该局政策法规司副司长吴丽雅说。《疫苗管理法》是全球首部综合性疫苗管理法律。具体来讲,吴丽雅介绍,生产、销售的疫苗属于假药的,没收违法所得和违法生产、销售的疫苗以及专门用于违法生产疫苗的原料、辅料、包装材料、设备等物品,责令停产停业整顿,吊销药品注册证书,直至吊销药品生产许可证等,并处违法生产、销售疫苗货值金额 15 倍以上 50 倍以下的罚款。生产、销售的疫苗属于劣药的,处违法生产、销售疫苗货款金额 10 倍以上 30 倍以下罚款。吴丽雅为读者算了一笔账。生产、销售的疫苗属于假药的,最低罚款:50 万元 ×(15 倍至 50 倍)=750 万元至 2500 万元;生产、销售的疫苗属于劣药的,最低罚款: 50 万元 ×(10 倍至 30 倍)=500 万元至 1500 万元。疫苗是特殊的药品,事关公共安全。吴丽雅表示,长春长生问题疫苗事件暴露出我国在疫苗监管方面存在主体责任不落实、质量安全管理不到位、职业化专业化监管力量薄弱等问题。鉴于此,《疫苗管理法》明确提出国家对疫苗实行最严格的管理制度,对疫苗的研制、生产、流通、...
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产品名称:

Premo™ FUCCI Cell Cycle Sensor (BacMam 2.0)

上市日期: 2019-04-18
产品类别: INVITROGEN

规格

Color:Green, Red-Orange
Delivery Method:Baculovirus (BacMam System)
Detection Method:Fluorescent
Emission Class:Visible
Excitation Class:Visible
Excitation⁄Emission (nm):488⁄510 (Green), 555⁄584 (Red)
For Use With (Equipment):Flow Cytometer, Fluorescence Microscope, Fluorescent Imager, High Content Instrument, Tali® Image-Based Cytometer
Form:Liquid
Label or Dye:GFP (EmGFP), RFP (TagRFP)
Product Line:Premo™
Product Size:2 x 200 µL
Quantity:2 vials (200 µl each)
Sub-Cellular Localization:Nucleus
Technique:Fluorescence Intensity
Shipping Condition:Wet Ice


  • 产品描述
  • 产品功能
  • 产品参数

描述

Premo™ FUCCI Cell Cycle Sensor is a fluorescent, two-color sensor of cell cycle progression and division in live cells. It allows accurate and sensitive cell cycle analysis of individual cells or a population of cells by fluorescence microscopy, flow cytometry, or high-content imaging.

At 2 x 200 µL, this is a smaller pack size version (1/5 volume) of the popular full-size product and perfect for testing its performance in your system.

The Premo™ FUCCI Cell Cycle Sensor is delivered by highly efficient BacMam 2.0 technology, enabling cell cycle studies in essentially any cell type. Provided in a ready-to-use format—simply add, incubate, and image. Premo™ FUCCI affords highly efficient transient expression in cell lines, primary cells, and stem cells.

Premo™ FUCCI Cell Cycle Sensor is:

Accurate: Cell-cycle controlled expression of bright GFP and RFP indicators for live cell analysis of individual cells or populations
Highly Efficient: >90% transduction of a wide range of mammalian cell lines, including primary cells, stem cells, and neurons
Fast and Convenient: Simply add Premo™ FUCCI Cell Cycle Sensor to your cells in complete medium, incubate overnight, and analyze
Safe: BacMam reagents are non-integrating and non-replicating in mammalian cells, lack observable cytopathic effects, and are suitable for biosafety level (BSL) 1 handling

Study Cell Cycle in the Relevant Cellular Context
If you are studying cell cycle regulators, cell differentiation, or are developing anti-cancer compounds against cell cycle-related targets, Premo™ FUCCI is a superbly sensitive and accurate sensor tool. The efficient transduction and transient expression allows you to conduct these experiments in the biologically relevant cell types.

Simple, Quick Cell Cycle Visualization
Simply add the ready-to-use Premo™ FUCCI cell cycle sensor to your cells in complete medium, incubate overnight, then visualize cell cycle progression in populations of cells using fluorescence microscopy, flow cytometry, or high content imaging instruments. Cells change from red in the G1 to yellow in the G1/S interphase and green in S, G2, and M phases, as fusions of emGFP and TagRFP coupled to two cell cycle-regulated proteins are expressed and degraded.

How It Works
Premo™ FUCCI is derived from the fluorescence ubiquitination cell cycle indicator (FUCCI), developed by Miyawaki and colleagues (Ref 1). FUCCI is based on two cell cycle-regulated proteins, geminin and Cdt1, fused to one green (emGFP) and red (TagRFP) fluorescent protein, respectively. As Cdt1 and geminin are present only during specific phases of the cell cycle, the fluorescent protein partner is similarly cell-cycle dependent. Ubiquitination by specific ubiquitin E3 ligases target the fusion constructs in the proteasome for degradation. E3 ligases display temporal regulation of activity, resulting in biphasic cycling of geminin and Cdt1 levels during the cell cycle. Geminin-GFP is degraded in the G1 phase; the presence of Cdt1-TagRFP is indicated by red fluorescence within nuclei. During the S, G2, and M phases, however, Cdt1 is degraded and only geminin-GFP remains, giving cells green-fluorescent nuclei. During the G1/S transition, when Cdt1 levels are decreasing and geminin levels are increasing, both proteins are present, giving a yellow-fluorescent nuclear signal. This dynamic color change (red to yellow to green) clearly displays progression through the cell cycle and division.

Powered by BacMam 2.0 Technology
BacMam technology is based on an insect virus (baculovirus) for efficient transduction and transient expression in mammalian cells. The Premo™ FUCCI cell cycle sensor uses BacMam 2.0 technology to enable cell cycle studies in essentially any cell type, from cell lines to stem cells by incorporation of a mammalian expression cassette for the FUCCI construct. Compared with BacMam 1.0 technology, BacMam 2.0 does not require the use of an enhancer and can be used in complete medium. There is no need to purify plasmid or worry about vector integrity and quality. No lipids, dye-loading chemicals, or other potentially harmful treatments are required. BacMam 2.0 incorporates elements that greatly enhance transduction efficiency and expression levels: a pseudotyped capsid protein for more efficient cell entry, an enhanced CMV promoter, and the Woodchuck hepatitis post-transcriptional regulatory element (WPRE).

Titratable Expression that Lasts Up to Two Weeks
The transient transgene expression lasts from about 5 days in transformed cell lines to up to two weeks in slowly dividing cells, such as some primary cell types; we have imaged terminally differentiated neurons more than three weeks after transduction. In addition, BacMam technology permits defined optimization because it gives you the ability to precisely titrate expression levels. Baculoviruses do not replicate in mammalian cells, nor are viral genes expressed, giving them an excellent safety profile and lack of cytopathic effects on mammalian cells.

Learn more about the Premo™ FUCCI Cell Cycle Sensor

Reference:

1. Sakaue-Sawano A, Kurokawa H, Morimura T, Hanyu A, Hama H, Osawa H, Kashiwagi S, Fukami K, Miyata T, Miyoshi H, Imamura T, Ogawa M, Masai H, Miyawaki A. Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. Cell. 2008 Feb 8;132(3):487-98. PubMed PMID: 18267078

Related products
We offer a range of BacMam-based reagents beyond CellLight® reagents, including the BacMam GFP Transduction Control that is ideal to test out the technology and optimize transduction conditions, Premo™ Biosensors, including Premo™ Autophagy Sensor, ion channel drug targets, pathway analysis kits, and more.

Learn more about these products and the BacMam technology

See other imaging tools and reagents

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use


规格

Color:Green, Red-Orange
Delivery Method:Baculovirus (BacMam System)
Detection Method:Fluorescent
Emission Class:Visible
Excitation Class:Visible
Excitation⁄Emission (nm):488⁄510 (Green), 555⁄584 (Red)
For Use With (Equipment):Flow Cytometer, Fluorescence Microscope, Fluorescent Imager, High Content Instrument, Tali® Image-Based Cytometer
Form:Liquid
Label or Dye:GFP (EmGFP), RFP (TagRFP)
Product Line:Premo™
Product Size:2 x 200 µL
Quantity:2 vials (200 µl each)
Sub-Cellular Localization:Nucleus
Technique:Fluorescence Intensity
Shipping Condition:Wet Ice


规格

Color:Green, Red-Orange
Delivery Method:Baculovirus (BacMam System)
Detection Method:Fluorescent
Emission Class:Visible
Excitation Class:Visible
Excitation⁄Emission (nm):488⁄510 (Green), 555⁄584 (Red)
For Use With (Equipment):Flow Cytometer, Fluorescence Microscope, Fluorescent Imager, High Content Instrument, Tali® Image-Based Cytometer
Form:Liquid
Label or Dye:GFP (EmGFP), RFP (TagRFP)
Product Line:Premo™
Product Size:2 x 200 µL
Quantity:2 vials (200 µl each)
Sub-Cellular Localization:Nucleus
Technique:Fluorescence Intensity
Shipping Condition:Wet Ice


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