EasySep™小鼠T细胞分选试剂盒

不带标记的小鼠T细胞免疫磁珠负选

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产品号 #（选择产品）

产品号 #19851_C

不带标记的小鼠T细胞免疫磁珠负选

产品优势

操作简单、快速
纯度高达99%
无需分离柱
获得不带标记的活细胞

产品组分包括

EasySep™小鼠T细胞分选试剂盒（产品号 #19854）

EasySep™小鼠T细胞分选抗体混合物，0.5 mL
EasySep™ Streptavidin RapidSpheres™ 50001磁珠，1 mL
EasySep™小鼠FcR阻断剂，0.2 mL

RoboSep™小鼠T细胞分选试剂盒（产品号 #19854RF）

EasySep™小鼠T细胞分选抗体混合物，0.5 mL
EasySep™ Streptavidin RapidSpheres™ 50001磁珠，1 mL
EasySep™小鼠FcR阻断剂，0.2 mL
RoboSep™ 缓冲液（产品号 #20104）
RoboSep™过滤吸头（产品号 #20125）

立即询价

总览

使用EasySep™小鼠T细胞分选试剂盒，通过免疫磁珠负选，可简便高效地从脾细胞或其他组织的单细胞悬液中分离高纯度的小鼠T细胞。EasySep™技术结合单克隆抗体的特异性和无柱磁分选系统的简便性，已在发表的研究中广泛应用超过20年。

在该EasySep™负选流程中，非目的细胞会被抗体复合物与磁珠标记。表达以下标志物的非目的细胞将被识别并去除：CD11b、CD45R、Ter119、CD49b、CD19和CD24。使用EasySep™磁极吸附后，通过简单地将目的细胞倾倒或吸取至一个新的试管中，即可将被磁珠标记的细胞与不带标记的目的细胞分离开来。在短至15分钟的磁珠分选后，目的T细胞可立即用于流式细胞术、培养及基于细胞的实验等下游应用。

了解更多EasySep™免疫磁珠技术的工作原理，或如何通过RoboSep™实现全自动化免疫磁珠细胞分选。探索为您的实验流程优化的更多产品，包括培养基、补充剂、抗体等。

实验数据

Figure 1.Typical EasySep™ Mouse T Cell Isolation Profile

Starting with mouse splenocytes, the T cell content (CD3+CD19-) of the isolated fraction is 96.6 ± 2.0% (mean ± SD), using the purple EasySep™ magnet.

Figure 2.Cell Isolation Protocol Lengths

Typical time taken (in minutes) to isolate cells using select EasySep™ kits.

Figure 3.ImmunoCult™ Mouse T Cell Activator Kit Supports High Viability of Activated T Cells

Mouse T cells were isolated using EasySep™ Mouse T Cell Isolation Kit (Catalog #19851), stimulated with ImmunoCult™ Mouse T Cell Activator Kit (Catalog #100-1572), and cultured in IMDM + FBS formulation. Following 3 days of culture, the mean ± SD frequency of CD25+ cells was 91.9 ± 5.1% (n = 11) or 99.9 ± 0.1% (n = 5), when stimulated with ImmunoCult™ Mouse CD3/CD28 T Cell Activator or ImmunoCult™ Mouse CD3/CD28/CD2 T Cell Activator, respectively. Stimulated mouse T cells maintained expression levels of CD25 throughout the 7-day culture period.

Figure 4.Robust Expansion of EasySep™-Isolated Mouse T Cells Can Be Achieved Following Stimulation with ImmunoCult™ Mouse T Cell Activator Kit

Mouse T cells isolated using EasySep™ Mouse T Cell Isolation Kit (Catalog #19851) were expanded with ImmunoCult™ Mouse T Cell Activator Kit (Catalog #100-1572) in IMDM + FBS formulation over 7 days. The number of viable cells was assessed every 2 - 3 days, and fresh medium supplemented with IL-2 was added. No additional ImmunoCult™ Mouse T Cell Activator was added during the 7-day culture period. After 7 days in culture with ImmunoCult™ Mouse CD3/CD28 T Cell Activator or ImmunoCult™ Mouse CD3/CD28/CD2 T Cell Activator, stimulation resulted in a fold expansion of 23 ± 3.4 or 29.3 ± 4.8 (mean ± SEM, n = 6), respectively.

Figure 5.High Cell Proliferation is Observed in EasySep™-Isolated T cells After Stimulation with ImmunoCult™ Mouse T Cell Activator

Mouse T cells isolated using EasySep™ Mouse T Cell Isolation Kit (Catalog #19851) were labeled with CFDA-SE (Catalog #75003), stimulated with ImmunoCult™ Mouse T Cell Activator Kit (Catalog #100-1572), and cultured in cultured in IMDM + FBS formulation. On Day 3, cells were harvested, stained with anti-mouse CD4 and CD8a antibodies, then measured by flow cytometry. Shown are CFDA-SE-labeled mouse T cells, gated on viable CD4+ (A) or CD8a+ (B) cells, cultured with no activator (top panel), with ImmunoCult™ Mouse CD3/CD28 T Cell Activator (middle panel), or with ImmunoCult™ Mouse CD3/CD28/CD2 T Cell Activator (bottom panel). Due to cell proliferation, the intensity of CFDA-SE signal is reduced by 50% for each cell division.

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明，或浏览下方更多实验方案。

应用领域

本产品专为以下研究领域设计，可应用于工作流程中的关键步骤（图中高亮）。探索这些工作流程，了解更多我们为各研究领域提供的其他配套产品。

常见问题

文献 (34)

S100a9 Protects Against the Effects of Repeated Social Defeat Stress. C. M. Moshfegh et al. Biological psychiatry global open science 2023 oct

Abstract

BACKGROUND Posttraumatic stress disorder, a consequence of psychological trauma, is associated with increased inflammation and an elevated risk of developing comorbid inflammatory diseases. However, the mechanistic link between this mental health disorder and inflammation remains elusive. We previously found that S100a8 and S100a9 messenger RNA, genes that encode the protein calprotectin, were significantly upregulated in T lymphocytes and positively correlated with inflammatory gene expression and the mitochondrial redox environment in these cells. Therefore, we hypothesized that genetic deletion of calprotectin would attenuate the inflammatory and redox phenotype displayed after psychological trauma. METHODS We used a preclinical mouse model of posttraumatic stress disorder known as repeated social defeat stress (RSDS) combined with pharmacological and genetic manipulation of S100a9 (which functionally eliminates calprotectin). A total of 186 animals (93 control, 93 RSDS) were used in these studies. RESULTS Unexpectedly, we observed worsening of behavioral pathology, inflammation, and the mitochondrial redox environment in mice after RSDS compared with wild-type animals. Furthermore, loss of calprotectin significantly enhanced the metabolic demand on T lymphocytes, suggesting that this protein may play an undescribed role in mitochondrial regulation. This was further supported by single-cell RNA sequencing analysis demonstrating that RSDS and loss of S100a9 primarily altered genes associated with mitochondrial function and oxidative phosphorylation. CONCLUSIONS These data demonstrate that the loss of calprotectin potentiates the RSDS-induced phenotype, which suggests that its observed upregulation after psychological trauma may provide previously unexplored protective functions.

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Co-modulation of T cells and B cells enhances the inhibition of inflammation in experimental hypersensitivity pneumonitis. O. Courtemanche et al. Respiratory research 2022 oct

Abstract

BACKGROUND Hypersensitivity pneumonitis (HP) is an interstitial lung disease characterized by antigen-triggered neutrophilic exacerbations. Although CD4+ T cells are sufficient for HP pathogenesis, this never translated into efficient T cell-specific therapies. Increasing evidence shows that B cells also play decisive roles in HP. Here, we aimed to further define the respective contributions of B and T cells in subacute experimental HP. METHODS Mice were subjected to a protocol of subacute exposure to the archaeon Methanosphaera stadmanae to induce experimental HP. Using models of adoptive transfers of B cells and T cells in Rag1-deficient mice and of B cell-specific S1P1 deletion, we assessed the importance of B cells in the development of HP by evaluating inflammation in bronchoalveolar lavage fluid. We also aimed to determine if injected antibodies targeting B and/or T cells could alleviate HP exacerbations using a therapeutic course of intervention. RESULTS Even though B cells are not sufficient to induce HP, they strongly potentiate CD4+ T cell-induced HP?‘associated neutrophilic inflammation in the airways. However, the reduction of 85% of lung B cells in mice with a CD19-driven S1P1 deletion does not dampen HP inflammation, suggesting that lung B cells are not necessary in large numbers to sustain local inflammation. Finally, we found that injecting antibodies targeting B cells after experimental HP was induced does not dampen neutrophilic exacerbation. Yet, injection of antibodies directed against B cells and T cells yielded a potent 76% inhibition of neutrophilic accumulation in the lungs. This inhibition occurred despite partial, sometimes mild, depletion of B cells and T cells subsets. CONCLUSIONS Although B cells are required for maximal inflammation in subacute experimental HP, partial reduction of B cells fails to reduce HP-associated inflammation by itself. However, co-modulation of T cells and B cells yields enhanced inhibition of HP exacerbation caused by an antigenic rechallenge.

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Immune correlates of protection following Rift Valley fever virus vaccination. J. D. Doyle et al. NPJ vaccines 2022 oct

Abstract

Rift Valley fever virus (RVFV) is a hemorrhagic fever virus with the potential for significant economic and public health impact. Vaccination with an attenuated strain, DelNSsRVFV, provides protection from an otherwise lethal RVFV challenge, but mechanistic determinants of protection are undefined. In this study, a murine model was used to assess the contributions of humoral and cellular immunity to DelNSsRVFV-mediated protection. Vaccinated mice depleted of T cells were protected against subsequent challenge, and passive transfer of immune serum from vaccinated animals to na{\{i}}ve animals was also protective demonstrating that T cells were dispensable in the presence of humoral immunity and that humoral immunity alone was sufficient. Animals depleted of B cells and then vaccinated were protected against challenge. Total splenocytes but not T cells alone B cells alone or B??+??T cells harvested from vaccinated animals and then transferred to na{\"{i}}ve animals were sufficient to confer protection suggesting that multiple cellular interactions were required for effective cellular immunity. Together these data indicate that humoral immunity is sufficient to confer vaccine-mediated protection and suggests that cellular immunity plays a role in protection that requires the interaction of various cellular components."

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更多信息

更多信息
物种	小鼠
Magnet Compatibility	• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000)
样本来源	其它细胞系, 脾脏
Selection Method	Negative

EasySep™小鼠T细胞分选试剂盒

产品号 #19851

产品号 #19851RF

产品号 #（选择产品）

产品号 #19851_C

产品优势

产品组分包括

总览

实验数据

产品说明书及文档

应用领域

相关材料与文献

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Scientists Helping Scientists™

EasySep™小鼠T细胞分选试剂盒

产品号 #19851

产品号 #19851RF

产品号 #（选择产品）

产品号 #19851_C

产品优势

产品组分包括

总览

实验数据

产品说明书及文档

应用领域

相关材料与文献

技术资料 (13)

常见问题

Can EasySep™ Streptavidin RapidSpheres™ be used for either positive or negative selection?

How does the separation work?

Which columns do I use?

How can I analyze the purity of my enriched sample?

Can EasySep™ Streptavidin RapidSphere™ separations be automated?

Are cells isolated using EasySep™ RapidSphere™ products FACS-compatible?

Can I alter the separation time in the magnet?

文献 (34)

Abstract

Abstract

Abstract

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