{{'Search' | translate}}
 

Glycine

甘氨酸(白色晶体或结晶粉末),Fisher BioReagents TM

Company: Thermo Fisher Scientific
Catalog#: BP381-5
Bio-protocol()
Company-protocol()
Other protocol()

Isolation, Culture, and Staining of Single Myofibers
Author:
Date:
2016-10-05
[Abstract]  Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be recapitulated ex vivo by isolation of single myofibers from skeletal muscles and culturing them under suspension conditions. Here, we describe an improved protocol to evaluate ex vivo satellite cells activation through isolation of single myofibers from extensor digitorum longus (EDL) muscle of mice and culturing and staining of myofiber-associated satellite cells with the markers of self-renewal, proliferation, and differentiation. [摘要]  成年骨骼肌再生由被称为卫星细胞的成人干细胞的专门群体协调,其被定位在基底层和肌纤维的质膜之间。在肌肉再生期间发生的卫星细胞激活,增殖和随后分化的过程可以通过从骨骼肌中分离单个肌纤维并在悬浮条件下培养它们而在体外重现。在这里,我们描述了一种改进的协议,通过从小鼠的伸肌腱长肌(EDL)肌肉中分离单个肌纤维来评价离体细胞的活化,并培养和染色肌纤维相关的卫星细胞的标记物自我更新,增殖和分化。

[背景] 虽然骨骼肌是一种完全分化的有丝分裂后组织,但是它保持了内在的能力,以对遗传和获得性肌肉纤维损伤的形式Grand和Rudnicki,2007)。成年人的肌肉再生由称为卫星细胞的干细胞群体介导,所述细胞位于有丝分裂静止状态的肌纤维的基底层和肌纤维膜之间(Le Grand和Rudnicki,2007)。响应于肌肉创伤,卫星细胞变得活化和增殖以产生与预先存在的纤维融合的成肌细胞,并彼此相互修复或产生新的肌纤维。一小部分卫星细胞不分化,而是重新进入静止以维持干细胞库。所有哺乳动物物种的卫星细胞表达配对盒(Pax)转录因子Pax7,其也用作确定与其他生肌因子如MyoD相关的卫星细胞命运的关键标记(Le Grand和Rudnicki,2007; et al。,2006; Kuang and Rudnicki,2008)。
  可以通过肌纤维外植体的悬浮培养部分地重现关于卫星细胞活化,增殖和分化的肌肉再生的体内过程(Rosenblatt等人,1995; ...

Monoclonal Antibody Purification (Nicotiana benthamiana Plants)
Author:
Date:
2014-01-20
[Abstract]  Plant-based expression systems provide an alternative biomanufacturing platform for recombinant proteins (Matoba et al., 2011). In particular, plant virus-based vectors can overexpress proteins within days in the leaf tissue of Nicotiana benthamiana (N. benthamiana). To overcome the issues of genetic instability and limited infectivity of recombinant viruses, Agrobacterium-mediated delivery of “deconstructed” virus vectors has become the mainstay for the production of large and/or multicomponent proteins, such as immunoglobulin (Ig)G monoclonal antibodies (mAbs). Here, we describe a method of producing human IgG mAbs in N. benthamiana using the tobamoviral replicon vector magnICON®. The vector can express up to a few hundred mg of a ... [摘要]  基于植物的表达系统为重组蛋白提供了替代的生物制造平台(Matoba等人,2011)。特别地,基于植物病毒的载体可以在烟草(Nicotiana benthamiana)(本塞姆氏烟草)的叶组织中天数内过表达蛋白质。为了克服重组病毒的遗传不稳定性和有限的感染性的问题,土壤杆菌介导的"解构的"病毒载体的递送已经成为生产大和/或多组分蛋白如免疫球蛋白的主要方法Ig)G单克隆抗体(mAb)。在这里,我们描述了在N中产生人IgG mAbs的方法。本生烟草使用烟草花叶病毒复制子载体magnICON 。载体在7天内可以表达高达几百mg的mAb/kg叶材料。显示了分别用于广泛中和的抗HIV和抗流感mAbs VRC01和CR6261的代表性病例(Hamorsky等人,2013)。叶组织匀浆,通过过滤和离心澄清提取物。 mAb通过快速蛋白质液相色谱(FPLC)使用蛋白A亲和力和Phenyl HP疏水界面树脂纯化。

Comments