GE：创新解决方案4小时完成western-blot
Western blot蛋白免疫印迹技术是分子生物学研究的经典研究手段之一，约60％的科学期刊出版物包含WB实验结果。但WB实验步骤多，耗时长，要获得一个完美的结果十分不易，WB实验结果的不稳定，重复不出来？定量不准确？凝胶又漏了？
GE公司最新推出的Amersham WB系统，4小时内可完成电泳、转印、杂交、孵育、成像、定量分析，一气呵成，为Western blot提供了整体解决方案。每个样品、每一次实验都能获得一致的定量数据。再也无需担心结果难以重复再现，无需反复摸索实验条件！
Protein synthesis: a high fidelity molecular event
Rachel Green (Johns Hopkins U., HHMI) 1: Protein synthesis: a high fidelity molecular event
Talk Overview:
In her first talk, Green provides a detailed look at protein synthesis, or translation. Translation is the process by which nucleotides, the “language” of DNA and RNA, are translated into amino acids, the “language” of proteins. Green begins by describing the components needed for translation; mRNA, tRNA, ribosomes, and the initiation, elongation, and termination factors. She then explains the roles of these players in ensuring accuracy during the initiation, elongation, termination and recycling steps of the translation process. By comparing translation in bacteria and eukaryotes, Green explains that it is possible to determine which components and steps are highly conserved and predate the divergence of different kingdoms on the tree of life, and which are more recent adaptations.
Green’s second talk focuses on work from her lab investigating how ribosomes detect defective mRNAs and trigger events leading to the degradation of the bad RNA and the incompletely translated protein product and to the recycling of the ribosome components. Working in yeast and using a number of biochemical and genetic techniques, Green’s lab showed that the protein Dom34 is critical for facilitating ribosome release from the short mRNAs that result from mRNA cleavage. Experiments showed that Dom34-mediated rescue of ribosomes from short mRNAs is an essential process for cell survival in higher eukaryotes.
Speaker Biography:
Rachel Green received her BS in chemistry from the University of Michigan. She then moved to Harvard to pursue her PhD in the lab of Jack Szostak where she worked on designing catalytic RNA molecules and investigating their implications for the evolution of life. As a post-doctoral fellow at the University of California, Santa Cruz, Green began to study how the ribosome translates mRNA to protein with such accuracy.
Currently, Green is a Professor of Molecular Biology and Genetics at the Johns Hopkins School of Medicine and an Investigator of the Howard Hughes Medical Institute. Research in her lab continues to focus on the ribosome and factors involved in the fidelity of eukaryotic and prokaryotic translation.
Green is the recipient of a Johns Hopkins University School of Medicine Graduate teaching Award as well as the recipient for numerous awards for her research. She was elected to the National Academy of Sciences in 2012.
Protein synthesis: mRNA surveillance by the ribosome
Rachel Green (Johns Hopkins U., HHMI) 2: Protein synthesis: mRNA surveillance by the ribosome
Talk Overview:
In her first talk, Green provides a detailed look at protein synthesis, or translation. Translation is the process by which nucleotides, the “language” of DNA and RNA, are translated into amino acids, the “language” of proteins. Green begins by describing the components needed for translation; mRNA, tRNA, ribosomes, and the initiation, elongation, and termination factors. She then explains the roles of these players in ensuring accuracy during the initiation, elongation, termination and recycling steps of the translation process. By comparing translation in bacteria and eukaryotes, Green explains that it is possible to determine which components and steps are highly conserved and predate the divergence of different kingdoms on the tree of life, and which are more recent adaptations.
Green’s second talk focuses on work from her lab investigating how ribosomes detect defective mRNAs and trigger events leading to the degradation of the bad RNA and the incompletely translated protein product and to the recycling of the ribosome components. Working in yeast and using a number of biochemical and genetic techniques, Green’s lab showed that the protein Dom34 is critical for facilitating ribosome release from the short mRNAs that result from mRNA cleavage. Experiments showed that Dom34-mediated rescue of ribosomes from short mRNAs is an essential process for cell survival in higher eukaryotes.
Speaker Biography:
Rachel Green received her BS in chemistry from the University of Michigan. She then moved to Harvard to pursue her PhD in the lab of Jack Szostak where she worked on designing catalytic RNA molecules and investigating their implications for the evolution of life. As a post-doctoral fellow at the University of California, Santa Cruz, Green began to study how the ribosome translates mRNA to protein with such accuracy.
Currently, Green is a Professor of Molecular Biology and Genetics at the Johns Hopkins School of Medicine and an Investigator of the Howard Hughes Medical Institute. Research in her lab continues to focus on the ribosome and factors involved in the fidelity of eukaryotic and prokaryotic translation.
Green is the recipient of a Johns Hopkins University School of Medicine Graduate teaching Award as well as the recipient for numerous awards for her research. She was elected to the National Academy of Sciences in 2012.
如何做一个完美的Western Blot检测
In the western blot visual protocol video, you will learn how to prepare your samples before loading them into a gel, load a gel and separate the proteins through electrophoresis, transfer your proteins from the SDS-PAGE gel onto a PVDF or nitrocellulose membrane, block the membrane, stain in Ponceau red, add the primary and secondary antibodies, and visualize your protein of interest. Additional help can be found in the support section of http://www.novusbio.com, through our live chat service, or by calling us directly to talk with our elite customer and technical service scientists.
Western Blot 第1阶段：样品制备
Novus Biologicals Visual Protocols: In phase 1 of the western blot procedure, you will learn how to prepare your samples before loading them into a gel. Here we isolate protein from cultured cells, quantify total protein concentrations with a BCA assay, add loading buffer to the sample, and heat the sample. Additional help can be found in the support section of http://www.novusbio.com, through our live chat service, or by calling us directly to talk with our elite customer and technical service scientists.
Western Blot 第2阶段：蛋白电泳(SDS-PAGE)
Novus Biologicals Visual Protocols: In phase 2 of the western blot procedure, you will learn how to load a gel and separate the proteins through electrophoresis, based upon protein weight. Additional help can be found in the support section of http://www.novusbio.com, through our live chat service, or by calling us directly to talk with our elite customer and technical service scientists.
Western Blot 第3阶段：膜转移
Novus Biologicals Visual Protocols: In phase 3 of the western blot procedure, you will learn how to transfer your proteins from the SDS-PAGE gel onto a PVDF or nitrocellulose membrane. Here we remove the gel from the cassette, and stack it in a sandwich comprised of a sponge, filter paper, the gel, membrane, filter paper, and sponge. The negatively charged proteins will then transfer onto the membrane toward the positive current. Additional help can be found in the support section of http://www.novusbio.com, through our live chat service, or by calling us directly to talk with our elite customer and technical service scientists.
Western Blot 第4阶段：免疫印迹法
Novus Biologicals Visual Protocols: In phase 4 of the western blot procedure, you will learn how to block the membrane, stain in with Ponceau red, and add the primary and secondary antibodies. Vigorous washing when indicated between these steps is essential for obtaining a clean blot. Additional help can be found in the support section of www.novusbio.com, through our live chat service, or by calling us directly to talk with our elite customer and technical service scientists.
Western Blot 第5阶段：检测
Novus Biologicals Visual Protocols: In phase 5 of the western blot procedure, you will learn how to visualize your protein of interest that was probed with specific antibodies in the previous step. Here we utilize the electrochemiluminescent (ECL) to produce light where our antibodies are bound. This light is collected by film or a camera for later analysis. Additional help can be found in the support section of http://www.novusbio.com, through our live chat service, or by calling us directly to talk with our elite customer and technical service scientists.
CRISPR/Cas9 & tetraOne：基因敲除/敲入鼠模型的快速构建技术
众所周知，基因工程小鼠模型已被广泛应用于生物医药研究，但模型小鼠的构建技术复杂、耗时长且花费高，让很多实验室望而却步。在本次讲座中，欧阳应斌博士（赛业生物技术副总裁、高级科学家）主要介绍基因敲除/敲入鼠模型的快速构建技术——CRISPR/Cas9基因编辑技术与tetraOne技术，同时会简述转基因技术（PiggyBac系统）和传统ES打靶技术，并重点讲解每种技术的优势、缺点及应用。