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CD14 clone M5E2 (PE-Cy7)

Company: BD
Catalog#: 557742
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Protocol for Isolation, Stimulation and Functional Profiling of Primary and iPSC-derived Human NK Cells
Author:
Date:
2020-12-05
[Abstract]  

Natural killer (NK) cells are innate immune cells, characterized by their cytotoxic capacity, and chemokine and cytokine secretion upon activation. Human NK cells are identified by CD56 expression. Circulating NK cells can be further subdivided into the CD56bright (~10%) and CD56dim NK cell subsets (~90%). NK cell-like cells can also be derived from human induced pluripotent stem cells (iPSC). To study the chemokine and cytokine secretion profile of the distinct heterogenous NK cell subsets, intracellular flow cytometry staining can be performed. However, this assay is challenging when the starting material is limited. Alternatively, NK cell subsets can be enriched, sorted, stimulated, and functionally profiled by measuring secreted effector molecules in the supernatant by Luminex. Here,

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[摘要]  [摘要]天然杀伤(NK)细胞是先天性免疫细胞,其特征在于其细胞毒性能力以及活化后的趋化因子和细胞因子分泌。人NK细胞通过CD56表达鉴定。循环的NK细胞可进一步细分为CD56亮(约10%)和CD56暗NK细胞亚群(约90%)。NK细胞样细胞也可以源自人诱导的多能干细胞(iPSC)。为了研究不同的异源NK细胞亚群的趋化因子和细胞因子分泌概况,可以进行细胞内流式细胞仪染色。然而,当起始原料有限时,该测定法具有挑战性。或者,可以通过Luminex测量上清液中分泌的效应子分子来富集,分选,刺激和功能性分析NK细胞亚群。在这里,我们提供了一种快速直接的方案,用于分离和刺激原代NK细胞或iPSC衍生的NK细胞样细胞,并随后检测分泌的细胞因子和趋化因子,这也适用于少量细胞。


[背景]自然杀伤(NK)细胞是先天免疫系统的一部分,提供第一线防御病毒感染和畸形。在人血中,可以基于CD56和CD16表达鉴定出两个不同的NK细胞群体:CD56明亮的CD16 +/-和CD56暗的CD16 + ...

Flow Cytometry Measurement of Glucocerebrosidase Activity in Human Monocytes
Author:
Date:
2020-04-05
[Abstract]  Glucocerebrosidase (GCase) is an important enzyme for the metabolism of glycolipids. GCase enzyme deficiency is implicated in human disease and the efficient measurement of GCase activity is important for evaluating the efficacy of therapeutics targeting this enzyme. Existing approaches to measure GCase activity include whole blood mass spectrometry-based assays, where an internal standard is used to measure the accumulation of ceramide following metabolism of the synthetic substrate C12-glucocerebroside, and the utilisation of fluorescent probes that bind active GCase and/or release fluorescent metabolites upon cleavage by GCase. Here, we describe the application of a fluorescence-activated cell sorter-based assay to efficiently quantitate GCase enzyme activity in the monocyte population ... [摘要]  

[摘要 ] 葡萄糖脑苷脂酶(GCase )是糖脂代谢的重要酶。GCase 酶缺乏症与人类疾病有关,GCase 活性的有效测量对于评估靶向该酶的治疗剂的功效至关重要。现有的测量GCase 活性的方法包括基于全血质谱的分析,其中使用内标测量合成底物C12-葡萄糖脑苷代谢后神经酰胺的积累,以及利用结合活性GCase 和// 的荧光探针的利用。或通过GCase 裂解后释放荧光代谢物 。在这里,我们描述了基于荧光激活细胞分选术的测定方法的应用,以有效地定量人类外周血单核细胞单核细胞群中的GCase 酶活性。细胞渗透性的GCase 衬底5-(Pentafluorobenzoylamino )荧光素二-β-D- 吡喃葡萄糖苷(PFB- FDGlu )提供了一个用于测量的GCase 活性,由此酶裂解产生的绿色荧光PFB-F染料,在FL-检测流式细胞仪的1个通道。使用溶酶体GCase 活性抑制剂,conduritol B-环氧,以确保特异性。该协议为测量活的单个细胞中的GCase 活性提供了一种有利的方法。

[背景 ] 葡萄糖脑苷脂酶(GCase ),由 GBA1基因是一种溶酶体水解酶,可将葡糖神经酰胺转化为葡萄糖和神经酰胺。GBA1 ...

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