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Company: BD

Catalog#: 552311

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A Simple and Robust Protocol for in vitro Differentiation of Mouse Non-pathogenic T Helper 17 Cells from CD4⁺ T Cells

Siwen Kang, Ruohan Wu and Ruoning Wang

Published online: 2021-05-20

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Intracellular Cytokine Staining (ICS) on Human Lymphocytes or Peripheral Blood Mononuclear Cells (PBMCs)

Author:

Sheena Gupta and Holden Maecker,

Date:

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。

Enrichment of Cells of Interest from Heterogeneous Murine Cells with BioMag Goat Anti-rat IgG

Author:

Hongcheng Wang,

Date:

2012-04-05

[Abstract] BioMag Goat Anti-Rat IgG is a standard BioMag particle coated with polyclonal goat anti-rat IgG antibodies and is highly suited for use in cell sorting methods where a rat IgG antibody is used as a primary antibody. BioMag Goat Anti-Rat IgG can be used to separate the cells of interest from a heterogeneous cell population using negative selection. BioMag Goat Anti-Rat IgG can also be used as a secondary antibody in enzyme immunoassays and radioassays that utilize a rat IgG primary monoclonal antibody.
The efficiency of enrichment by negative selection depends on antigen availability and the total cell population. The author has intensive experience about enrichment of mouse CD4CD8 double negative (DN) thymocytes from total thymocytes which contains only about 2% DN population. ... [摘要] BioMag Goat抗大鼠IgG是用多克隆山羊抗大鼠IgG抗体包被的标准BioMag颗粒，并且非常适用于其中将大鼠IgG抗体用作第一抗体的细胞分选方法。 BioMag山羊抗大鼠IgG可用于使用阴性选择从异质细胞群体中分离感兴趣的细胞。 BioMag山羊抗大鼠IgG也可用作酶免疫测定和利用大鼠IgG第一单克隆抗体的放射性测定中的第二抗体。通过阴性选择的富集效率取决于抗原可用性和总细胞群体。作者有丰富的丰富的小鼠CD4CD8双阴性（DN）胸腺细胞从总胸腺细胞，只含有约2％的DN人口。通常在一轮富集后，DN人群占总胸腺细胞的80％以上。然而，骨髓细胞的消耗效率低得多。

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