EasySep™ Release人CD45正选试剂盒

采用磁珠解离技术对人CD45+白细胞（包括TILs）进行免疫磁珠正选

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产品号 #（选择产品）

产品号 #100-0105_C

采用磁珠解离技术对人CD45+白细胞（包括TILs）进行免疫磁珠正选

产品优势

在45分钟内分离出高纯度的人CD45+细胞
无需清洗去除EasySep™ Releasable RapidSpheres™可解离磁珠

产品组分包括

EasySep™ Release人CD45正选试剂盒（产品号 #100-0105）

EasySep™ Release人CD45正选抗体混合物，0.5 mL
EasySep™ Releasable RapidSpheres™磁珠，1 mL
EasySep™ Release缓冲液（浓缩），6 x 1 mL

RoboSep™ Release人CD45正选试剂盒（产品号 #100-0108）

EasySep™ Release人CD45正选抗体混合物，0.5 mL
EasySep™ Releasable RapidSpheres™磁珠，1 mL
EasySep™ Release缓冲液（浓缩），6 x 1 mL
RoboSep™ 缓冲液（产品号 #20104）
RoboSep™过滤吸头（产品号 #20125）
EasySep™ EasyTube™-14（产品号 #20128）

用于人源化小鼠的EasySep™ Release 人CD45正选试剂盒（产品号 #100-0107）

EasySep™ Release人CD45正选抗体混合物，0.5 mL
EasySep™ Releasable RapidSpheres™磁珠，1 mL
EasySep™ Release缓冲液（浓缩），6 x 1 mL
EasySep™ 小鼠FcR阻断剂，0.5 mL

用于人源化小鼠的RoboSep™ Release 人CD45正选试剂盒（产品号 #100-0109）

EasySep™ Release人CD45正选抗体混合物，0.5 mL
EasySep™ Releasable RapidSpheres™磁珠，1 mL
EasySep™ Release缓冲液（浓缩），6 x 1 mL
EasySep™ 小鼠FcR阻断剂，0.5 mL
RoboSep™ 缓冲液（产品号 #20104）
RoboSep™ 过滤吸头（产品号 #20125）x 2
EasySep™ EasyTube™-14（产品号 #20128）× 2

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总览

使用EasySep™ Release人CD45正选试剂盒，可通过免疫磁珠正选，轻松从人原代组织和肿瘤样本（产品号 #100-0105）或人源化小鼠的组织和肿瘤异种移植物（产品号 #100-0107）的单细胞悬液中分离高纯度且无磁珠的人CD45+白细胞。EasySep™结合了单克隆抗体的特异性和无柱磁性系统的简便性，迄今已广泛应用于发表的研究中超过20年。

在此EasySep™正选过程中，目的细胞被识别CD45的抗体复合物和EasySep™ Releasable RapidSpheres™可解离磁珠标记。与常规的结合目的细胞的磁珠不同，这种磁珠具有可解离性。使用EasySep™磁极进行分选后，通过解离试剂去除经EasySep™分选的CD45+细胞上结合的磁珠。最终分选获得的高纯度CD45+细胞可直接用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。使用该EasySep™ Release试剂盒分选之后，细胞表面仍结合有抗体复合物，并可能与Brilliant Violet™偶联的抗体、聚乙二醇修饰的蛋白质或其他化学相关配体相互作用。

了解更多关于免疫磁珠EasySep™技术的工作原理，或如何通过RoboSep™实现全自动化免疫磁珠细胞分选。探索更多优化您实验流程的产品，包括培养基、添加剂、抗体等配套试剂。

实验数据

$Using the EasySep™ Release Human CD45 Positive Selection Kit, the CD45+ TIL purities of the start and final isolated fractions are 5.5% and 96.0%, respectively.$

Figure 1. EasySep™ Release Human CD45 Positive Selection Kit

Starting with a single-cell suspension of a human breast cancer tumor xenograft (MDA-MB-231) sample from an NRG-3GS humanized mouse, the CD45+ TIL purities of the start and final isolated fractions are 5.5% and 96.0%, respectively.
NOTE: Cell debris and dead cells were excluded from the analysis based on DRAQ5™ and DAPI fluorescence.

Using PBMCs as a startnig sample, this t-SNE plot shows that EasySep™-isolated CD45+ cells are representative of the starting leukocyte population.

Figure 2. EasySep™-Isolated CD45+ Cells are Representative of the Starting Leukocyte Population

Mass cytometry data comparing the composition of immune subsets in PBMCs and EasySep™-isolated cells from the same donor. Starting with whole blood, PBMCs were prepared by density gradient centrifugation using Lymphoprep™. To compare immune subset composition pre- and post-EasySep™ isolation, a fraction of the PBMCs was further isolated using EasySep™ Release Human CD45 Positive Selection Kit and the pre- and post-isolated fractions were assessed by mass cytometry (CyTOF®). t-SNE plots of cells stained with 19 markers and analyzed by CyTOF® are shown (n = 1).

PBMCs pre- and post-EasySep™ isolation were incubated in peptide pools (CEF, CPI, or PHA) and a robust population of IFN-gamma-producing cells in these respective samples are visible on ELISpot plates, showing that EasySep™-isolated CD45+ cells produce IFN-gamma in response to antigen and mitogen stimulation.

Figure 3. EasySep™-Isolated CD45+ Cells Produce IFN-gamma in Response to Antigen and Mitogen Stimulation

PBMCs pre- and post-EasySep™ isolation were incubated for 24 hours in the presence of peptide pools (CEF for antigen-specific CD8+ T cell response and CPI for antigen-specific CD4+ T cell response) or mitogen (phytohemagglutinin [PHA]). Following incubation, ELISpot plates were processed and IFN-gamma-producing cells were counted using an AID ELISpot reader. Representative images of ELISpot assays are shown (n = 3).

CD45+ cells isolated from spleens, lungs, bone marrow, MDA MB231 tumors, and SKOV3 tumors using EasySep™ are highly purified.

Figure 4. CD45+ Cells Isolated by EasySep™ from Various Tissues Are Highly Purified

A humanized mouse tumor model was generated by engraftment of human CD34+ hematopoietic stem and progenitor cells into NRG-3GS mice followed by xenotransplantation with human cancer cell lines, MDA-MB-231 (breast cancer) or SKOV-3 (ovarian carcinoma). Starting with a single-cell suspension of spleens, lungs, bone marrow, or tumors, human CD45+ leukocytes were isolated using EasySep™ Release Human CD45 Positive Selection Kit. The starting frequencies and isolated purities of human CD45+ cells of individual experiments and averages are shown.

文献 (1)

Dynamic alterations in PD-1/PD-L1 expression level and immune cell profiles based on radiation response status in mouse tumor model. Y. N. Yoon et al. Frontiers in oncology 2022

Abstract

INTRODUCTION Based on the immunologic effects of anti-cancer treatment and their therapeutic implications, we evaluated radiotherapy (RT)-induced dynamic alterations in programmed death-1 (PD-1)/PD ligand-1 (PD-L1) expression profiles. METHODS Local RT with 2 Gy ?— 5 or 7.5 Gy ?— 1 was administered to the CT26 mouse model. Thereafter, tumors were resected and evaluated at the following predefined timepoints according to radiation response status: baseline, early (immediately after RT), middle (beginning of tumor shrinkage), late (stable status with RT effect), and progression (tumor regrowth). PD-1/PD-L1 activity and related immune cell profiles were quantitatively assessed. RESULTS RT upregulated PD-L1 expression in tumor cells from the middle to late phase; however, the levels subsequently decreased to levels comparable to baseline in the progression phase. RT with 2 Gy ?— 5 induced a higher frequency of PD-L1+ myeloid-derived suppressor cells, with a lesser degree of tumor regression, compared to 7.5 Gy. The proportion of PD-1+ and interferon (IFN)-$\gamma$+CD8$\alpha$ T cells continued to increase. The frequency of splenic PD-1+CD8+ T cells was markedly elevated, and was sustained longer with 2 Gy ?— 5. Based on the transcriptomic data, RT stimulated the transcription of immune-related genes, leading to sequentially altered patterns. DISCUSSION The dynamic alterations in PD-1/PD-L1 expression level were observed according to the time phases of tumor regression. This study suggests the influence of tumor cell killing and radiation dosing strategy on the tumor immune microenvironment.

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种属	Human, Mouse
Magnet Compatibility	• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • EasyEights™ EasySep™ Magnet (Catalog #18103) • EasyPlate™ EasySep™ Magnet (Catalog #18102) • RoboSep™-S (Catalog #21000)
样本来源	Other, Tumor
Selection Method	Positive

EasySep™ Release人CD45正选试剂盒

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产品号 #100-0105_C

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EasySep™ Release人CD45正选试剂盒

产品号 #（选择产品）

产品号 #100-0105_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

相关材料与文献

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文献 (1)

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