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Poly-L-lysine solution

聚-L-赖氨酸溶液

Company: Sigma-Aldrich
Catalog#: P8920
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Preparation of Cerebellum Granule Neurons from Mouse or Rat Pups and Evaluation of Clostridial Neurotoxin Activity and Their Inhibitors by Western Blot and Immunohistochemistry
Author:
Date:
2018-07-05
[Abstract]  Cerebellar Granule Neurons (CGN) from post-natal rodents have been widely used as a model to study neuronal development, physiology and pathology. CGN cultured in vitro maintain the same features displayed in vivo by mature cerebellar granule cells, including the development of a dense neuritic network, neuronal activity, neurotransmitter release and the expression of neuronal protein markers. Moreover, CGN represent a convenient model for the study of Clostridial Neurotoxins (CNT), most notably known as Tetanus and Botulinum neurotoxins, as they abundantly express both CNT receptors and intraneuronal substrates, i.e., Soluble N-ethylmaleimide-sensitive factor activating protein receptors (SNARE proteins). Here, we describe a protocol for obtaining a highly pure ... [摘要]  来自产后啮齿动物的小脑颗粒神经元(CGN)已被广泛用作研究神经元发育,生理学和病理学的模型。 CGN体外培养维持成熟小脑颗粒细胞在体内显示的相同特征,包括发育致密的神经炎网络,神经元活动,神经递质释放和神经元的表达 蛋白质标记。 此外,CGN代表了梭菌神经毒素(CNT)研究的便利模型,最着名的是破伤风和肉毒杆菌神经毒素,因为它们大量表达CNT受体和神经元内基质, ie ,可溶性N-乙基马来酰亚胺 - 敏感因子激活蛋白受体(SNARE蛋白)。 在这里,我们描述了从出生后大鼠/小鼠获得高纯度CGN培养物的方案和用CNT中毒的简便方法。 我们还说明了评估CNT活性及其抑制的方便方法。

【背景】梭菌神经毒素(CNT)的大家族由破伤风神经毒素(TeNT)和肉毒杆菌神经毒素(BoNT)的多种变体形成,它们分别是破伤风和肉毒中毒的神经麻痹毒素(Schiavo et al。,2000; Johnson和Montecucco,2008; Rossetto et al。,2014)。 TeNT,七种BoNT血清型(BoNT / A至/ G)及其许多亚型是金属蛋白酶,通过切割SNARE蛋白(可溶性N-乙基马来酰亚胺敏感因子激活蛋白受体),三种必需蛋白质来阻断神经递质的释放而引起神经麻痹。控制突触小泡与突触前质膜的融合(Rossetto et al。,2014; ...

Imaging Cytokine Concentration Fields Using PlaneView Imaging Devices
Author:
Date:
2018-04-05
[Abstract]  We describe here a method to visualize concentration fields of cytokines around cytokine-secreting cells. The main challenge is that physiological cytokine concentrations can be very low, in the pico-molar range. Since it is currently impossible to measure such concentrations directly, we rely on cell’s response to the cytokines–the phosphorylation of a transcription factor–that can be visualized through antibody staining. Our devices aim at mimicking conditions in dense tissues, such as lymph nodes. A small number of secreting cells is deposited on a polylysine-coated glass and covered by multiple layers of cytokine-consuming. The cells are left to communicate for 1 h, after which the top layers are removed and the bottom layer of cells is antibody labeled for the response to cytokines. ... [摘要]  我们在这里描述了一种可视化细胞因子分泌细胞周围细胞因子浓度场的方法。主要挑战是生理细胞因子浓度可能非常低,在微摩尔浓度范围内。由于目前不可能直接测量这样的浓度,我们依赖于细胞对细胞因子的反应 - 转录因子的磷酸化 - 可以通过抗体染色显现。我们的设备旨在模仿密集组织中的条件,如淋巴结。少数分泌细胞沉积在聚赖氨酸包被的玻璃上并被多层细胞因子消耗覆盖。将细胞连通1小时,之后去除顶层,并且细胞的底层被抗体标记为对细胞因子的应答。然后通过标准荧光显微镜观察细胞因子场的横截面。这篇手稿总结了我们的方法,以量化密集细胞体外细胞因子介导的细胞间通讯的程度。

【背景】哺乳动物的免疫系统已经发展到能够识别和限制潜在病原体的传播,同时使由免疫系统本身造成的附带组织损伤最小化。为了实现这一点,免疫细胞依赖细胞因子介质网络,这些细胞因子介质能够进行细胞间通讯并广播关于致病性侮辱的大小和性质的信息。大量不同细胞因子与其同源受体强烈结合,通常在纳摩尔或皮摩尔范围内具有特征性结合亲和力。通过细胞因子通讯产生免疫龛。例如,在骨髓和胸腺中,通过基质细胞分泌的白细胞介素-7(IL-7)分别支持增殖的B细胞和T细胞祖细胞的存活(Tokoyoda et al。, 2004; Alves等人,2009)。细胞因子生态位的大小控制成熟祖细胞的数量,从而保持血细胞区室平衡(Böyum,1968; ...

Histochemical Staining of Collagen and Identification of Its Subtypes by Picrosirius Red Dye in Mouse Reproductive Tissues
Author:
Date:
2017-11-05
[Abstract]  Collagen is one of the foremost components of tissue extracellular matrix (ECM). It provides strength, elasticity and architecture to the tissue enabling it to bear the wear and tear from external factors like physical stress as well as internal stress factors like inflammation or other pathological conditions. During normal pregnancy or pregnancy related pathological conditions like preterm premature rupture of membranes (PPROM), collagen of the fetal membrane undergoes dynamic remodeling defining biochemical properties of the fetal membrane. The protocol in this article describes the histochemical method to stain total collagen by Picrosirius red stain which is a simple, quick and reliable method. This protocol can be used on paraformaldehyde (PFA) and formaldehyde fixed paraffin ... [摘要]  胶原蛋白是组织细胞外基质(ECM)最重要的组成部分之一。它为组织提供了强度,弹性和构造,使其能承受来自外部因素的磨损,如身体压力以及炎症或其他病理状况等内部应激因素。在正常妊娠或与妊娠相关的病理状况如胎膜早破(PPROM)期间,胎膜的胶原经历动态重塑,定义胎膜的生化特性。本文中的方案描述了一种简单,快速,可靠的组织化学染色方法,用苦皮红染色法对总胶原进行染色。该协议可用于多聚甲醛(PFA)和甲醛固定石蜡包埋组织切片。我们进一步描述了胶原蛋白在不同的小鼠生殖组织中的染色和分布,并且还展示了这种技术如何与偏振显微镜组合用于检测不同亚型胶原蛋白的分布。

【背景】胶原蛋白是从皮肤到骨头的所有结缔组织中的主要负载聚合物。当施加机械力时,胶原蛋白网络强烈变硬,从而防止组织的过度变形。有16种胶原蛋白,其中I型,II型和III型胶原蛋白几乎占据了人体胶原蛋白的80%,这些胶原蛋白被挤在一起形成细长的原纤维。 IV型胶原蛋白形成二维网状结构;而其他几种胶原蛋白类型与原纤维型胶原蛋白相关,将它们彼此连接或连接到其他基质组分。这些胶原蛋白与细胞外基质(ECM)的其他成分一起经历不断的重塑以提供所需的生物化学性质如拉伸强度和弹性。这种胶原蛋白的独特属性是生殖组织稳定性的影响因素之一,其失调可能导致不良事件,例如胎盘异常,膜破裂(Hampson等人,1997; Marpaung, ...

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