{{'Search' | translate}}



Company: Sigma-Aldrich
Catalog#: S0389
Other protocol()

Measuring CD38 Hydrolase and Cyclase Activities: 1,N6-Ethenonicotinamide Adenine Dinucleotide (ε-NAD) and Nicotinamide Guanine Dinucleotide (NGD) Fluorescence-based Methods
[Abstract]  CD38 is a multifunctional enzyme involved in calcium signaling and Nicotinamide Adenine Dinucleotide (NAD+) metabolism. Through its major activity, the hydrolysis of NAD+, CD38 helps maintain the appropriate levels of this molecule for all NAD+-dependent metabolic processes to occur. Due to current advances and studies relating NAD+ decline and the development of multiple age-related conditions and diseases, CD38 gained importance in both basic science and clinical settings. The discovery and development of strategies to modulate its function and, possibly, treat diseases and improve health span put CD38 under the spotlights. Therefore, a consistent and reliable method to measure its activity and explore its use in medicine is required. We ... [摘要]  CD38是参与钙信号传导和烟酰胺腺嘌呤二核苷酸(NAD + )代谢的多功能酶。通过其主要活性,NAD + 的水解,CD38有助于维持该分子的适当水平,从而发生所有NAD + 依赖性代谢过程。由于目前有关NAD + 下降和多种年龄相关疾病和疾病发展的研究和进展,CD38在基础科学和临床环境中都具有重要意义。调整其功能,可能治疗疾病和改善健康状况的战略的发现和发展使CD38成为焦点。因此,需要一种一致且可靠的方法来测量其活性并探索其在医学中的用途。我们在这里描述了我们的小组如何测量CD38的水解酶和环化酶活性的方法,利用基于荧光的酶测定法在平板阅读器中使用1,N 6 -Ethenonicotinamide Adenine Dinucleotide(ε-NAD)进行)和烟酰胺鸟嘌呤二核苷酸(NGD)分别作为底物。

【背景】目前关于代谢功能障碍和虚弱的年龄相关发展的研究每天都有更多的证据。众所周知,随着老化的进行,NAD + 水平在预期过程中降低。最近的研究表明,烟酰胺腺嘌呤二核苷酸(NAD + )的减少与年龄相关的代谢下降有关(Massudi et al。,2012)。增加NAD + 水平体内,导致延长寿命和健康跨度相关因素的活化,并改善衰老的几个生理和代谢参数(Camacho-Pereira ...

Separation of Thylakoid Protein Complexes with Two-dimensional Native-PAGE
[Abstract]  The hierarchical composition and interactions of the labile thylakoid protein complexes can be assessed by sequential 2D-native gel-electrophoresis system. Mild non-ionic detergent digitonin is used to solubilize labile protein super-and megacomplexes, which are then separated with first-dimension blue native polyacrylamide gel electrophoresis (1D-BN-PAGE). The digitonin derived protein complexes are further solubilized with stronger detergent, β-DM, and subsequently separated on an orthogonal 2D-BN-PAGE to release smaller protein subcomplexes from the higher-order supercomplexes. Here we describe a detailed method for 2D-BN-PAGE analysis of thylakoid protein complexes from Arabidopsis thaliana. [摘要]  不稳定的类囊体蛋白复合物的分级组成和相互作用可以通过连续的2D天然凝胶电泳系统来评估。 温和的非离子洗涤剂洋地黄皂苷用于溶解不稳定的蛋白质超级和巨型复合物,然后用第一维蓝色天然聚丙烯酰胺凝胶电泳(1D-BN-PAGE)分离。 将洋地黄皂苷衍生的蛋白质复合物用更强的去污剂β-DM进一步溶解,随后在正交的2D-BN-PAGE上分离,以从较高级的超复合物中释放较小的蛋白质亚复合物。 在这里,我们描述了来自拟南芥的类囊体蛋白复合物的2D-BN-PAGE分析的详细方法。

【背景】在类囊体膜中发生光合作用的光反应,在高等植物中,由贴壁的grana类囊体和非贴壁的基质类囊体组成。光反应由多亚基蛋白复合物光系统(PS)I和II,细胞色素b 6 f和ATP酶催化。 PSII及其光捕获天线复合物(LHCII)在grana-thylakoids中最为丰富,因此在空间上与基质类囊体定位的PSI-LHCI复合物隔离(Andersson和Anderson,1980)。格拉纳和基质类囊体之间的间期在两个光系统中都得到了丰富(Albertsson,2001; Suorsa et al。,2015)。通过光依赖性LHCII和PSII蛋白的可逆磷酸化介导,光系统与LHCII一起组装成更大的超级和超级复合物。 ...

Mouse Satellite Cell Isolation and Transplantation
[Abstract]  Satellite cell (SC) transplantation represents a powerful strategy to investigate SC biology during muscle regeneration. We described here a protocol for SC isolation from green fluorescent protein (GFP)-expressing mice and their transplantation into murine muscles. This procedure was originally used to assess the effects of the hormone unacylated ghrelin on muscle regeneration, in particular evaluating how the increase of unacylated ghrelin in the recipient muscle affected the engraftment of donor SCs (Reano et al., 2017). [摘要]  卫星细胞(SC)移植代表了肌肉再生期间SC生物学研究的强大策略。 我们在这里描述了从绿色荧光蛋白(GFP)表达的小鼠和他们的小鼠肌肉移植SC分离的协议。 该程序最初用于评估激素非酰化生长素释放肽对肌肉再生的影响,特别是评估受体肌肉中未酰化的生长素释放肽的增加如何影响供体SC的植入(Reano等人,2017年)。


由于移植的成肌细胞可以与宿主成肌细胞融合,因此SC的移植被认为是杜氏肌营养不良症(DMD)的潜在疗法多年的时间,这提示了在缺陷纤维中功能性修复的可能性(Partridge ...