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PerCP-CyTM5.5 Mouse Anti-Human CD4  Clone  SK3

PerCP-CyTM5.5 Mouse Anti-Human CD4  Clone  SK3

Company: BD
Catalog#: 341654
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Intracellular Cytokine Staining (ICS) on Human Lymphocytes or Peripheral Blood Mononuclear Cells (PBMCs)
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2015-04-05
[Abstract]  Production of cytokines plays an important role in the immune response. Cytokines are involved in many different pathways including the induction of many anti-viral proteins by IFN gamma, the induction of T cell proliferation by IL-2 and the inhibition of viral gene expression and replication by TNF alpha. Cytokines are not preformed factors but are rapidly produced and secreted in response to cellular activation. Intracellular cytokine detection by flow cytometry has emerged as the premier technique for studying cytokine production at the single-cell level. It detects the production and accumulation of cytokines within the endoplasmic reticulum after cell stimulation, allowing direct TH1 versus TH2 determination. It can also be used in combination with other flow cytometry protocols for ... [摘要]  细胞因子的产生在免疫应答中起重要作用。细胞因子参与许多不同的途径,包括通过IFNγ诱导许多抗病毒蛋白,通过IL-2诱导T细胞增殖以及抑制病毒基因表达和TNFα的复制。细胞因子不是预先形成的因子,但是响应于细胞激活而快速产生和分泌。通过流式细胞术的细胞内细胞因子检测已经成为研究单细胞水平的细胞因子产生的首要技术。它检测细胞刺激后细胞因子在内质网内的产生和积累,允许直接TH1对TH2测定。它也可以与其他流式细胞术协议结合使用细胞表面标记或与MHC多聚体进行免疫分型,以检测抗原特异性反应,使其成为一种非常灵活和通用的方法。这种能力,结合仪器的高通量性质,使细胞内细胞因子染色与现有的单细胞技术如ELISPOT,有限稀释和T细胞克隆相比具有巨大的优势。细胞内细胞因子染色的主要步骤如下:
1。使用特异性肽或非特异性活化混合物将细胞活化几小时;
2。加入蛋白质转运抑制剂(例如布雷菲德菌素A)以将细胞因子保留在细胞内;
。接下来,加入EDTA以从活化容器中除去粘附细胞;
4。洗涤后,可将细胞表面标记的抗体加入细胞中;
5。然后将细胞固定在多聚甲醛中并透化;
6。加入抗细胞因子抗体,并且可以通过流式细胞仪分析细胞。

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