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EasySep™人单核细胞富集试剂盒（不去除CD16）

免疫磁珠负选未标记的人CD14+CD16+单核细胞
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产品号 #（选择产品）

产品号 #19058_C

免疫磁珠负选未标记的人CD14+CD16+单核细胞

产品优势

  • 操作简单、快捷，且无需分离柱
  • 纯度高达90%
  • 获得的活细胞无标记

产品组分包括

  • EasySep™人单核细胞富集试剂盒（不去除CD16），（产品号 #19058）
    • EasySep™人单核细胞富集抗体混合物（不去除CD16）， 1 mL
    • EasySep™ 磁珠，1 mL
  • EasySep™人单核细胞富集试剂盒（不去除CD16），（产品号 #100-1525）
    • EasySep™人单核细胞富集抗体混合物（不去除CD16）， 1 x 10 mL
    • EasySep™ 磁珠，1 x 10 mL
  • RoboSep™ 人单核细胞富集试剂盒（不去除CD16，含过滤吸头），（产品号19058RF）
    • EasySep™人单核细胞富集抗体混合物（不去除CD16）， 1 mL
    • EasySep™ 磁珠，1 mL
    • RoboSep™ 缓冲液（产品号 #20104）
    • RoboSep™ 过滤吸头（产品号 #20125）
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New look, same high quality and support! You may notice that your instrument or reagent packaging looks slightly different from images displayed on the website, or from previous orders. We are updating our look but rest assured, the products themselves and how you should use them have not changed. Learn more
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  1. EasySep™ Magnet
    EasySep™ Magnet

    Magnet for column-free immunomagnetic separation

  2. EasySep™ Buffer
    EasySep™ Buffer

    Cell separation buffer

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
相关产品

总览

使用EasySep™ 人单核细胞富集试剂盒（不去除CD16），通过免疫磁珠负选，可轻松高效地从新鲜或冻存的人外周血单个核细胞（PBMCs）或裂解的白细胞单采术样本中分离高纯度的人CD14+CD16+单核细胞。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，非目的细胞被抗体复合物与磁珠标记。以下非目的细胞将被特异性去除：粒细胞、T细胞、B细胞、部分树突状细胞亚群、NK细胞及红系细胞。抗体混合物中还含有人Fc受体抗体，可最大程度地减少非特异性结合。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的细胞分离，接着简单地将目的细胞倾倒或吸取至一个新的试管中。经磁珠分选后，所得单核细胞及CD14+CD16+单核细胞即可用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。CD14+CD16+单核细胞亚群（约占健康人外周血的10%）具有组织巨噬细胞特性，在急慢性炎症性疾病中会显著扩增。

若需去除所有CD16+细胞，推荐使用含抗CD16抗体的EasySep™ 人单核细胞富集试剂盒（产品号 #19359）。

如需从裂解的白细胞单采术样本中大规模分选CD14+CD16+/-和CD14-CD16+CD56-单核细胞，请选用大规格试剂盒（1x10^10 个细胞，产品号 #100-1525）。

了解更多关于免疫磁珠EasySep™技术的工作原理，或如何通过RoboSep™实现免疫磁珠细胞分选全自动化。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。

在该EasySep™负选流程中，非目的细胞被抗体复合物与磁珠标记。以下非目的细胞将被特异性去除：粒细胞、T细胞、B细胞、部分树突状细胞亚群、NK细胞及红系细胞。抗体混合物中还含有人Fc受体抗体，可最大程度地减少非特异性结合。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的细胞分离，接着简单地将目的细胞倾倒或吸取至一个新的试管中。经磁珠分选后，所得单核细胞及CD14+CD16+单核细胞即可用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。CD14+CD16+单核细胞亚群（约占健康人外周血的10%）具有组织巨噬细胞特性，在急慢性炎症性疾病中会显著扩增。

若需去除所有CD16+细胞，推荐使用含抗CD16抗体的EasySep™ 人单核细胞富集试剂盒（产品号 #19359）。

如需从裂解的白细胞单采术样本中大规模分选CD14+CD16+/-和CD14-CD16+CD56-单核细胞，请选用大规格试剂盒（1x10^10 个细胞，产品号 #100-1525）。

了解更多关于免疫磁珠EasySep™技术的工作原理，或如何通过RoboSep™实现免疫磁珠细胞分选全自动化。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。

在该EasySep™负选流程中，非目的细胞被抗体复合物与磁珠标记。以下非目的细胞将被特异性去除：粒细胞、T细胞、B细胞、部分树突状细胞亚群、NK细胞及红系细胞。抗体混合物中还含有人Fc受体抗体，可最大程度地减少非特异性结合。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的细胞分离，接着简单地将目的细胞倾倒或吸取至一个新的试管中。经磁珠分选后，所得单核细胞及CD14+CD16+单核细胞即可用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。CD14+CD16+单核细胞亚群（约占健康人外周血的10%）具有组织巨噬细胞特性，在急慢性炎症性疾病中会显著扩增。

若需去除所有CD16+细胞，推荐使用含抗CD16抗体的EasySep™ 人单核细胞富集试剂盒（产品号 #19359）。

如需从裂解的白细胞单采术样本中大规模分选CD14+CD16+/-和CD14-CD16+CD56-单核细胞，请选用大规格试剂盒（1x10^10 个细胞，产品号 #100-1525）。

了解更多关于免疫磁珠EasySep™技术的工作原理，或如何通过RoboSep™实现免疫磁珠细胞分选全自动化。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。

在该EasySep™负选流程中，非目的细胞被抗体复合物与磁珠标记。以下非目的细胞将被特异性去除：粒细胞、T细胞、B细胞、部分树突状细胞亚群、NK细胞及红系细胞。抗体混合物中还含有人Fc受体抗体，可最大程度地减少非特异性结合。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的细胞分离，接着简单地将目的细胞倾倒或吸取至一个新的试管中。经磁珠分选后，所得单核细胞及CD14+CD16+单核细胞即可用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。CD14+CD16+单核细胞亚群（约占健康人外周血的10%）具有组织巨噬细胞特性，在急慢性炎症性疾病中会显著扩增。

若需去除所有CD16+细胞，推荐使用含抗CD16抗体的EasySep™ 人单核细胞富集试剂盒（产品号 #19359）。

如需从裂解的白细胞单采术样本中大规模分选CD14+CD16+/-和CD14-CD16+CD56-单核细胞，请选用大规格试剂盒（1x10^10 个细胞，产品号 #100-1525）。

了解更多关于免疫磁珠EasySep™技术的工作原理，或如何通过RoboSep™实现免疫磁珠细胞分选全自动化。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。

磁体兼容性
• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyPlate™ EasySep™ Magnet (Catalog 18102) • RoboSep™-S (Catalog #21000) • Easy 250 EasySep™ Magnet (Catalog #100-0821)
 
亚型
细胞分选试剂盒
 
细胞类型
单核细胞
 
种属
人
 
样本来源
PBMC
 
筛选方法
Negative
 
应用
细胞分选
 
品牌
EasySep，RoboSep
 
研究领域
免疫
 

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实验数据

FACS Histogram Results Using EasySep™ Human Monocyte Enrichment Kit Without CD16 Depletion

Figure 1. FACS Histogram Results Using EasySep™ Human Monocyte Enrichment Kit Without CD16 Depletion

Starting with freshly prepared PBMCs, the CD14+ cell content of the enriched fraction is 82.7 ± 6.7% (mean ± SD using the purple EasySep™ magnet). Slightly lower CD14+ cell purities may be obtained from samples that contain a large number of CD16+ cells. In the above example, the start and final enriched fractions are 21.2% and 83.3%, respectively.

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。

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet
Product Name
EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion
Catalog #
100-1525
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ Human Monocyte Enrichment Kit without CD16 Depletion with Filter Tips
Catalog #
19058RF
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion
Catalog #
19058
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human Monocyte Enrichment Kit without CD16 Depletion with Filter Tips
Catalog #
19058RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human Monocyte Enrichment Kit without CD16 Depletion with Filter Tips
Catalog #
19058RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human Monocyte Enrichment Kit without CD16 Depletion with Filter Tips
Catalog #
19058RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion
Catalog #
19058
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion
Catalog #
19058
Lot #
All
Language
English

应用领域

本产品专为以下研究领域设计，适用于工作流程中的高亮阶段。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。

Research Area
Workflow Stages

相关材料与文献

技术资料 (9)

Human Monocyte Research Product Workflow
Brochure
Human Monocyte Research Product Workflow
Tools to Streamline your Macrophage Research
Brochure
Tools to Streamline your Macrophage Research
EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Antigen Processing and Presentation
Wallchart
Antigen Processing and Presentation
Human Immune Cytokines
Wallchart
Human Immune Cytokines
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
1:57
Video
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
1:37
Video
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep...
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
1:13
Video
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology

常见问题

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

Yes. The EasySep™ kits use either a negative selection approach by targeting and removing unwanted cells or a positive selection approach targeting desired cells. Depletion kits are also available for the removal of cells with a specific undesired marker (e.g. GlyA).

How does the separation work?

Magnetic particles are crosslinked to cells using Tetrameric Antibody Complexes (TAC). When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a separate fraction.

Which columns do I use?

The EasySep™ procedure is column-free. That's right - no columns!

How can I analyze the purity of my enriched sample?

The Product Information Sheet provided with each EasySep™ kit contains detailed staining information.

Can EasySep™ separations be automated?

Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.

Can EasySep™ be used to isolate rare cells?

Yes. We recommend a cell concentration of 2x108 cells/mL and a minimum working volume of 100 µL. Samples containing 2x107 cells or fewer should be suspended in 100 µL of buffer.

Are the EasySep™ magnetic particles FACS-compatible?

Yes, the EasySep™ particles are flow cytometry-compatible, as they are very uniform in size and about 5000X smaller than other commercially available magnetic beads used with column-free systems.

Can the EasySep™ magnetic particles be removed after enrichment?

No, but due to the small size of these particles, they will not interfere with downstream applications.

Can I alter the separation time in the magnet?

Yes; however, this may impact the kit's performance. The provided EasySep™ protocols have already been optimized to balance purity, recovery and time spent on the isolation.

For positive selection, can I perform more than 3 separations to increase purity?

Yes, the purity of targeted cells will increase with additional rounds of separations; however, cell recovery will decrease.

How does the binding of the EasySep™ magnetic particle affect the cells? is the function of positively selected cells altered by the bound particles?

Hundreds of publications have used cells selected with EasySep™ positive selection kits for functional studies. Our in-house experiments also confirm that selected cells are not functionally altered by the EasySep™ magnetic particles.

If particle binding is a key concern, we offer two options for negative selection. The EasySep™ negative selection kits can isolate untouched cells with comparable purities, while RosetteSep™ can isolate untouched cells directly from whole blood without using particles or magnets.

文献 (22)

Interleukin-4 receptor alpha signaling regulates monocyte homeostasis. P. Haider et al. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2022 oct

Abstract

Interleukin-4 (IL-4) and its receptors (IL-4R) promote the proliferation and polarization of macrophages. However, it is unknown if IL-4R also influences monocyte homeostasis and if steady state IL-4 levels are sufficient to affect monocytes. Employing full IL-4 receptor alpha knockout mice (IL-4R$\alpha$-/- ) and mice with a myeloid-specific deletion of IL-4R$\alpha$ (IL-4R$\alpha$f/f LysMcre ), we show that IL-4 acts as a homeostatic factor regulating circulating monocyte numbers. In the absence of IL-4R$\alpha$, murine monocytes in blood were reduced by 50% without altering monocytopoiesis in the bone marrow. This reduction was accompanied by a decrease in monocyte-derived inflammatory cytokines in the plasma. RNA sequencing analysis and immunohistochemical staining of splenic monocytes revealed changes in mRNA and protein levels of anti-apoptotic factors including BIRC6 in IL-4R$\alpha$-/- knockout animals. Furthermore, assessment of monocyte lifespan in vivo measuring BrdU+ cells revealed that the lifespan of circulating monocytes was reduced by 55% in IL-4R$\alpha$-/- mice, whereas subcutaneously applied IL-4 prolonged it by 75%. Treatment of human monocytes with IL-4 reduced the amount of dying monocytes in vitro. Furthermore, IL-4 stimulation reduced the phosphorylation of proteins involved in the apoptosis pathway, including the phosphorylation of the NF$\kappa$Bp65 protein. In a cohort of human patients, serum IL-4 levels were significantly associated with monocyte counts. In a sterile peritonitis model, reduced monocyte counts resulted in an attenuated recruitment of monocytes upon inflammatory stimulation in IL-4R$\alpha$f/f LysMcre mice without changes in overall migratory function. Thus, we identified a homeostatic role of IL-4R$\alpha$ in regulating the lifespan of monocytes in vivo.
View publication
BCG hydrogel promotes CTSS-mediated antigen processing and presentation, thereby suppressing metastasis and prolonging survival in melanoma. M. Kremenovic et al. Journal for immunotherapy of cancer 2022 jun

Abstract

BACKGROUND The use of intralesional Mycobacterium bovis BCG (intralesional live BCG) for the treatment of metastatic melanoma resulted in regression of directly injected, and occasionally of distal lesions. However, intralesional-BCG is less effective in patients with visceral metastases and did not significantly improve overall survival. METHODS We generated a novel BCG lysate and developed it into a thermosensitive PLGA-PEG-PLGA hydrogel (BCG hydrogel), which was injected adjacent to the tumor to assess its antitumor effect in syngeneic tumor models (B16F10, MC38). The effect of BCG hydrogel treatment on contralateral tumors, lung metastases, and survival was assessed to evaluate systemic long-term efficacy. Gene expression profiles of tumor-infiltrating immune cells and of tumor-draining lymph nodes from BCG hydrogel-treated mice were analyzed by single-cell RNA sequencing (scRNA-seq) and CD8+ T cell receptor (TCR) repertoire diversity was assessed by TCR-sequencing. To confirm the mechanistic findings, RNA-seq data of biopsies obtained from in-transit cutaneous metastases of patients with melanoma who had received intralesional-BCG therapy were analyzed. RESULTS Here, we show that BCG lysate exhibits enhanced antitumor efficacy compared to live mycobacteria and promotes a proinflammatory tumor microenvironment and M1 macrophage (M$\Phi$) polarization in vivo. The underlying mechanisms of BCG lysate-mediated tumor immunity are dependent on M$\Phi$ and dendritic cells (DCs). BCG hydrogel treatment induced systemic immunity in melanoma-bearing mice with suppression of lung metastases and improved survival. Furthermore, BCG hydrogel promoted cathepsin S (CTSS) activity in M$\Phi$ and DCs, resulting in enhanced antigen processing and presentation of tumor-associated antigens. Finally, BCG hydrogel treatment was associated with increased frequencies of melanoma-reactive CD8+ T cells. In human patients with melanoma, intralesional-BCG treatment was associated with enhanced M1 M$\Phi$, mature DC, antigen processing and presentation, as well as with increased CTSS expression which positively correlated with patient survival. CONCLUSIONS These findings provide mechanistic insights as well as rationale for the clinical translation of BCG hydrogel as cancer immunotherapy to overcome the current limitations of immunotherapies for the treatment of patients with melanoma.
View publication
Activation of the Innate Immune Checkpoint CLEC5A on Myeloid Cells in the Absence of Danger Signals Modulates Macrophages' Function but Does Not Trigger the Adaptive T Cell Immune Response. M. J. Tosiek et al. Journal of immunology research 2022

Abstract

C-Type lectin receptor 5A (CLEC5A) is a spleen tyrosine kinase- (Syk-) coupled pattern recognition receptor expressed on myeloid cells and involved in the innate immune response to viral and bacterial infections. Activation of the CLEC5A receptor with pathogen-derived antigens leads to a secretion of proinflammatory mediators such as TNF-$\alpha$ and IL-6 that may provoke a systemic cytokine storm, and CLEC5A gene polymorphisms are associated with the severity of DV infection. In addition, the CLEC5A receptor was mentioned in the context of noninfectious disorders like chronic obstructive pulmonary disease (COPD) or arthritis. Altogether, CLEC5A may be considered as an innate immune checkpoint capable to amplify proinflammatory signals, and this way contributes to infection or to aseptic inflammation. In this study, we determined CLEC5A receptor expression on different macrophage subsets (in vitro and ex vivo) and the functional consequences of its activation in aseptic conditions. The CLEC5A surface expression appeared the highest on proinflammatory M1 macrophages while intermediate on tumor-associated phenotypes (M2c or TAM). In contrast, the CLEC5A expression on ex vivo-derived alveolar macrophages from healthy donors or macrophages from ovarian cancer patients was hardly detectable. Targeting CLEC5A on noninflammatory macrophages with an agonistic $\alpha$-CLEC5A antibody triggered a release of proinflammatory cytokines, resembling a response to dengue virus, and led to phenotypic changes in myeloid cells that may suggest their reprogramming towards a proinflammatory phenotype, e.g., upregulation of CD80 and downregulation of CD163. Interestingly, the CLEC5A agonist upregulated immune-regulatory molecules like CD206, PD-L1, and cytokines like IL-10, macrophage-derived chemokine (MDC/CCL22), and thymus and activation chemokine (TARC/CCL17) which are associated with an anti-inflammatory or a protumorigenic macrophage phenotype. In the absence of concomitant pathogenic or endogenous danger signals, the CLEC5A receptor activation did not amplify an autologous T cell response, which may represent a protective innate mechanism to avoid an undesirable autoimmune adaptive response.
View publication
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更多信息

更多信息
种属 Human
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyPlate™ EasySep™ Magnet (Catalog 18102) • RoboSep™-S (Catalog #21000) • Easy 250 EasySep™ Magnet (Catalog #100-0821)
样本来源 PBMC
Selection Method Negative
标记抗体
  1. 抗人CD16抗体，克隆号3G8
    抗人CD16抗体，克隆号3G8

    小鼠抗人、恒河猴、食蟹猴CD16单克隆IgG1抗体

  2. 抗人CD14抗体，克隆M5E2
    抗人CD14抗体，克隆M5E2

    抗人、恒河猴、食蟹猴CD14的小鼠单克隆IgG2a抗体

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