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EasySep™ Direct 人 PBMC 分选试剂盒

直接从全血中免疫磁珠负选不带标记的人外周血单个核细胞
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产品号 #19654_C

直接从全血中免疫磁珠负选不带标记的人外周血单个核细胞

产品优势

  • 9%的红细胞去除率，无需密度梯度离心、沉降或裂解
  • 操作简单、快捷，且无需分离柱
  • 分选得到的细胞不带标记

产品组分包括

  • EasySep™ Direct 人 PBMC 分选试剂盒（产品号 #19654）
    • EasySep™ Direct 人 PBMC 分选抗体混合物，2 x 2.5 mL
    • EasySep™ Direct RapidSpheres™ 磁珠，4 x 2.5 mL
  • RoboSep™ Direct 人 PBMC分选试剂盒（产品号 #19654RF）
    • EasySep™ Direct 人 PBMC 分选抗体混合物，2 x 2.5 mL
    • EasySep™ Direct RapidSpheres™ 磁珠，4 x 2.5 mL
    • RoboSep™ 缓冲液（产品号 #20104）
    • RoboSep™ 过滤吸头（产品号 #20125）
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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™ Direct人PBMC分选试剂盒，可通过免疫磁珠负选技术简便高效地从新鲜全血、buffy coat、骨髓、脐带血或白细胞单采术样本中获得高纯度的人外周血单个核细胞（PBMCs）。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和 EasySep™ Direct RapidSpheres™ 磁珠标记。以下非目的细胞将被清除：粒细胞、血小板和红细胞。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。完成磁珠分选后，目的PBMCs 可立即用于流式细胞术、培养或DNA/RNA提取等下游应用。

进一步了解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以节省时间并提升实验室通量。探索更多为您的实验流程优化的产品，包括细胞鉴定、冻存等相关产品。

磁极兼容性
• EasySep™磁极（产品号 #18000）
• “The Big Easy” EasySep™磁极（产品号 #18001）
• EasyEights™ EasySep™磁极（产品号 #18103）
• Easy 50 EasySep™磁极（产品号 #18002）
 
分类
细胞分选试剂盒
 
细胞类型
B 细胞，淋巴细胞，单核细胞，单个核细胞，NK 细胞，T 细胞
 
种属
人
 
样本来源
骨髓、白膜层、脐带血、白细胞单采术样本、全血
 
分选方法
负选
 
品牌
EasySep，RoboSep
 
研究领域
免疫
 

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实验数据

Typical EasySep™ Direct Protocol

Figure 1. Typical EasySep™ Direct Protocol

Typical EasySep™ Direct Human PBMC Isolation Profile

Figure 2. Typical EasySep™ Direct Human PBMC Isolation Profile

In the above example, the mononuclear cell content of the whole blood start sample (lysed by ammonium chloride) and non-lysed final isolated fraction is 27.0% and 98.6% (not gated on CD45), respectively.

Representative Flow Cytometry Plots

Figure 3. Representative Flow Cytometry Plots

Representative Forward Scatter (FSC) vs. Side Scatter (SSC) flow cytometry plots of mononuclear cells isolated from whole blood samples using either density gradient centrifugation or EasySep™ Direct Human PBMC Isolation Kit.

Representative t-SNE plots of Isolated PBMCs

Figure 4. Representative t-SNE plots of Isolated PBMCs

Representative t-SNE plots of PBMCs stained with 19 markers and analyzed with mass cytometry (CyTOF). Cells are clustered and colored based on the combination of markers they express. Both density gradient centrifugation and EasySep™ Direct Human PBMC Isolation Kit resulted in comparable cell populations with the exception of the contaminating granulocyte population (CD16+CD15+CD11b+CD66b+ CD45low).

EasySep™ Direct Human PBMC Isolation Kit Results in Fewer Contaminating Cells Compared to Density Gradient Centrifugation

Figure 5. EasySep™ Direct Human PBMC Isolation Kit Results in Fewer Contaminating Cells Compared to Density Gradient Centrifugation

Mononuclear cells were isolated from whole blood samples using either density gradient centrifugation or EasySep™ Direct Human PBMC Isolation Kit. Cells were counted and analyzed by flow cytometry. (A) Both density gradient centrifugation and EasySep™ Direct Human PBMC Isolation Kit resulted in an equivalent total number of nucleated cells recovered from 24-hour blood samples (mean ± SD; n=14). (B) Using EasySep™ Direct Human PBMC Isolation Kit to obtain mononuclear cells from 24-hour old blood samples resulted in lower numbers of residual platelets (CD41+), red blood cells (Glycophorin A+/CD45-), and granulocytes (CD66b+) compared to density gradient centrifugation (mean ± SD; n=15). (C) Cell isolation from 24-hour, 48-hour, 72-hour and 96-hour old blood samples using EasySep™ Direct Human PBMC Isolation Kit resulted in lower numbers of residual granulocytes compared to cell isolation using density gradient centrifugation (mean ± SD; n=3).

PBMCs Isolated with EasySep™ Direct Human PBMC Isolation Kit Proliferate and Maintain High RNA Integrity

Figure 6. PBMCs Isolated with EasySep™ Direct Human PBMC Isolation Kit Proliferate and Maintain High RNA Integrity

Mononuclear cells were isolated from whole blood samples using either density gradient centrifugation or EasySep™ Direct Human PBMC Isolation Kit. (A) Isolated mononuclear cells were used for downstream RNA isolation. There was no significant difference in RNA integrity as measured with the Agilent RNA Bioanalyzer (mean ± SEM, n=3). (B) Isolated mononuclear cells were labeled with Proliferation Dye eFluor 450 and stimulated with ImmunoCult™ Human CD3/CD28 T Cell Activator and 0.5ng/ml IL-2. After 4 days in culture, cells were analyzed for proliferation by flow cytometry. Representative histogram showing dividing cells (eFluor 450low).

PBMC Isolation from a Full-Size Leukapheresis Pack Using the EasySep™ Direct Human PBMC Isolation Kit Automated with RoboSep™-S Results in Fewer Contaminating Cells Compared to Density Gradient Centrifugation

Figure 7. PBMC Isolation from a Full-Size Leukapheresis Pack Using the EasySep™ Direct Human PBMC Isolation Kit Automated with RoboSep™-S Results in Fewer Contaminating Cells Compared to Density Gradient Centrifugation

Mononuclear cells were isolated from single concentrated leukapheresis packs using either density gradient centrifugation with Lymphoprep™ density gradient medium (Density Gradient Centrifugation) or EasySep™ Direct Human PBMC Isolation Kit automated on the RoboSep™-S instrument (EasySep™ Direct on RoboSep™-S). Cells were counted and analyzed by flow cytometry. Compared to density gradient centrifugation, EasySep™ Direct Human PBMC Isolation Kit automated on the RoboSep™-S instrument resulted in (A) equivalent numbers of total mononuclear cells, (B) equivalent numbers of total monocytes and lymphocytes and (C) lower numbers of residual platelets (CD41+), red blood cells (Glycophorin A+ /CD45− ), and granulocytes (CD66b+ ) (mean ± SD n=6).

Easy 250 EasySep™ Magnet Can Be Used to Isolate Mononuclear Cells from Whole Blood and Leukopak Samples of Up to 125 mL

Figure 8. Easy 250 EasySep™ Magnet Can Be Used to Isolate Mononuclear Cells from Whole Blood and Leukopak Samples of Up to 125 mL

Mononuclear cells (MNCs) were isolated using the EasySep™ Direct Human PBMC Isolation Kit (Catalog #19654) from large whole blood samples (25 - 125 mL) or unprocessed leukopaks (25 - 125 mL) with the Easy 250 EasySep™ Magnet (Easy 250; Catalog #100-0821) and from small samples (1 mL whole blood) with the EasySep™ Magnet (Purple; Catalog #18000). (A) Representative flow cytometry plots show the MNC fraction (Glycophorin A- CD42b-). In the above example, the mononuclear cell content of the whole blood start sample (lysed by ammonium chloride) and non-lysed final isolated fraction is 28.9% and 97.1% (not gated on CD45), respectively. (B) Cell purity was measured before (Start) and after isolation (Easy 250, Purple) based on viable cells. Cells were counted and analyzed by flow cytometry. Data shown as mean ± SD; n = 7 - 9.

EasySep™ Direct Human PBMC Isolation Kit Reduces Platelet Contamination More Efficiently Than Column-Based Methods

Figure 9. EasySep™ Direct Human PBMC Isolation Kit Reduces Platelet Contamination More Efficiently Than Column-Based Methods

Peripheral blood mononuclear cells (PBMCs) were isolated from 24-hour-old whole blood samples using either the EasySep™ Direct Human PBMC Isolation Kit or alternative, column-based kits. Flow cytometry analysis demonstrated that PBMCs isolated with the EasySep™ kit contained significantly fewer contaminating platelets (CD42b+), while levels of residual red blood cells (Glycophorin A+/CD45-) and granulocytes (CD66b+) were comparable between methods. Data are presented as mean ± SD (n ≥ 3 per condition).

EasySep™ Direct PBMC Isolation Kit Achieves Higher Recovery and Purity of Mononuclear Cells Compared to Column-Based Kits

Figure 10. EasySep™ Direct PBMC Isolation Kit Achieves Higher Recovery and Purity of Mononuclear Cells Compared to Column-Based Kits

PBMCs were isolated from 24-hour-old whole blood samples using the EasySep™ Direct Human PBMC Isolation Kit and two column-based kits designed for PBMC isolation from whole blood. Isolated cells were labeled with anti-CD45 antibodies and analyzed by flow cytometry. Cell purity was assessed by the proportion of CD45+ events among total recorded events (serving as an indicator of debris), and total recovery was quantified across isolation methods. The EasySep™ Direct PBMC Isolation Kit yielded higher purity PBMCs than both Column-Based Kits A and B. It also recovered a greater number of CD45+ mononuclear cells compared to Column-Based Kit A, with a comparable recovery to Column-Based Kit B. Data are presented as mean ± SD (n ≥ 3 per condition).

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 - Buffy Coat
Product Name
RoboSep™ Direct Human PBMC Isolation Kit
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Product Information Sheet - Whole Blood
Product Name
RoboSep™ Direct Human PBMC Isolation Kit
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Product Information Sheet - Bone Marrow
Product Name
RoboSep™ Direct Human PBMC Isolation Kit
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Product Information Sheet - Leukoreduction System Chamber
Product Name
RoboSep™ Direct Human PBMC Isolation Kit
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Product Information Sheet - Leukapheresis
Product Name
RoboSep™ Direct Human PBMC Isolation Kit
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Product Information Sheet - Cord Blood
Product Name
RoboSep™ Direct Human PBMC Isolation Kit
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Product Information Sheet - Cord Blood
Product Name
EasySep™ Direct Human PBMC Isolation Kit
Catalog #
19654
Lot #
All
Language
English
Document Type
Product Information Sheet - Buffy Coat
Product Name
EasySep™ Direct Human PBMC Isolation Kit
Catalog #
19654
Lot #
All
Language
English
Document Type
Product Information Sheet - Whole Blood
Product Name
EasySep™ Direct Human PBMC Isolation Kit
Catalog #
19654
Lot #
All
Language
English
Document Type
Product Information Sheet - Bone Marrow
Product Name
EasySep™ Direct Human PBMC Isolation Kit
Catalog #
19654
Lot #
All
Language
English
Document Type
Product Information Sheet - Leukoreduction System Chamber
Product Name
EasySep™ Direct Human PBMC Isolation Kit
Catalog #
19654
Lot #
All
Language
English
Document Type
Product Information Sheet - Leukapheresis
Product Name
EasySep™ Direct Human PBMC Isolation Kit
Catalog #
19654
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Direct Human PBMC Isolation Kit
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Direct Human PBMC Isolation Kit
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Direct Human PBMC Isolation Kit
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Direct Human PBMC Isolation Kit
Catalog #
19654
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Direct Human PBMC Isolation Kit
Catalog #
19654
Lot #
All
Language
English

相关材料与文献

技术资料 (20)

EasySep™ Direct Human PBMC Isolation Kit
Brochure
EasySep™ Direct Human PBMC Isolation Kit
Cell Separation Products for HLA Analysis
Brochure
Cell Separation Products for HLA Analysis
EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Cell Separation Solutions for the Flow Cytometric Crossmatch Assay
Technical Bulletin
Cell Separation Solutions for the Flow Cytometric Crossmatch Assay
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
Human Immune Cytokines
Wallchart
Human Immune Cytokines
How to Isolate Cells Directly from Whole Blood - EasySep™ Direct Protocol, EasyEights Magnet
6:13
Video
How to Isolate Cells Directly from Whole Blood - EasySep™ Direct Protocol, EasyEights Magnet
How to Isolate Cells Directly from Whole Blood Using the EasySep™ Purple/Silver Magnets
4:30
Video
How to Isolate Cells Directly from Whole Blood Using the EasySep™ Purple/Silver Magnets
How to Prepare a Buffy Coat from Whole Blood
3:46
Video
How to Prepare a Buffy Coat from Whole Blood
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
How to Recover Cells from a Leukocyte Reduction System (LRS) Cone
4:55
Video
How to Recover Cells from a Leukocyte Reduction System (LRS) Cone
Cell Isolation Directly from Whole Blood without RBC Lysis or Centrifugation: EasySep™ Direct
1:44
Video
Cell Isolation Directly from Whole Blood without RBC Lysis or Centrifugation: EasySep™ Direct
How to Isolate Cells Directly from Whole Blood Using the Easy 50 EasySep™ Magnet: 30-Minute Protocol
5:17
Video
How to Isolate Cells Directly from Whole Blood Using the Easy 50 EasySep™ Magnet: 30-Minute Protoc...
How to Thaw Frozen Human Primary Cells
6:05
Video
How to Thaw Frozen Human Primary Cells
Optimize the Flow Cytometry Crossmatch Assay: Save Time Without Compromising Sensitivity
1:01:28
Webinar
Optimize the Flow Cytometry Crossmatch Assay: Save Time Without Compromising Sensitivity
SARS-CoV-2 Viral Diversity: Causes, Consequences, and Opportunities
45:33
Webinar
SARS-CoV-2 Viral Diversity: Causes, Consequences, and Opportunities
Cell Separation Solutions for HLA and Chimerism Analysis
26:43
Webinar
Cell Separation Solutions for HLA and Chimerism Analysis
Streamlining PBMC Isolation with Automation: A Biobank’s Validation Experience
51:35
Webinar
Streamlining PBMC Isolation with Automation: A Biobank’s Validation Experience
Centrifugation and RBC Lysis-Free Preparation of Blood Samples in Under 30 Minutes
Scientific Poster
Centrifugation and RBC Lysis-Free Preparation of Blood Samples in Under 30 Minutes
Human Immune Cell Isolation Course
On-Demand Training
Human Immune Cell Isolation Course

文献 (6)

An additional dose of viral vector COVID-19 vaccine and mRNA COVID-19 vaccine in kidney transplant recipients: A randomized controlled trial (CVIM 4 study). J. Bruminhent et al. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons 2022 nov

Abstract

Immunogenicity following an additional dose of Coronavirus disease 2019 (COVID-19) vaccine was investigated in an extended primary series among kidney transplant (KT) recipients. Eighty-five KT participants were randomized to receive either an mRNA (M group; n =??43) or viral vector (V group; n =??42) vaccine. Among them, 62% were male, with a median (IQR) age of 50 (43-59) years and post-transplantation duration of 46 (26-82) months. At 2??weeks post-additional dose, there was no difference in the seroconversion rate between the M and V groups (70% vs. 65%, p =??.63). A median (IQR) of anti-RBD antibody level was not statistically different between the M group compared with the V group (51.8 [5.1-591] vs. 28.5 [2.9-119.3] BAU/ml, p =??.18). Furthermore, the percentage of participants with positive SARS-CoV-2 surrogate virus neutralization test results was not statistically different between groups (20% vs. 15%, p =??.40). S1-specific T cell and RBD-specific B cell responses were also comparable between the M and V groups (230 [41-420] vs. 268 [118-510], p =??.65 and 2 [0-10] vs. 2 [0-13] spot-forming units/106 peripheral blood mononuclear cells, p =??.60). In conclusion, compared with an additional dose of viral vector COVID-19 vaccine, a dose of mRNA COVID-19 vaccine did not elicit significantly different responses in KT recipients, regarding either humoral or cell-mediated immunity. (TCTR20211102003).
View publication
SARS-CoV-2-specific humoral and cell-mediated immune responses after immunization with inactivated COVID-19 vaccine in kidney transplant recipients (CVIM 1 study). J. Bruminhent et al. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons 2022 mar

Abstract

Immunogenicity following inactivated SARS-CoV-2 vaccination among solid organ transplant recipients has not been assessed. Seventy-five patients (37 kidney transplant [KT] recipients and 38 healthy controls) received two doses, at 4-week intervals, of an inactivated whole-virus SARS-CoV-2 vaccine. SARS-CoV-2-specific humoral (HMI) and cell-mediated immunity (CMI) were measured before, 4 weeks post-first dose, and 2 weeks post-second dose. The median (IQR) age of KT recipients was 50 (42-54) years and 89% were receiving calcineurin inhibitors/mycophenolate/corticosteroid regimens. The median (IQR) time since transplant was 4.5 (2-9.5) years. Among 35 KT patients, the median (IQR) of anti-RBD IgG level measured by CLIA after vaccination was not different from baseline, but was significantly lower than in controls (2.4 [1.1-3.7] vs. 1742.0 [747.7-3783.0] AU/ml, p < .01) as well as percentages of neutralizing antibody inhibition measured by surrogate viral neutralization test (0 [0-0] vs. 71.2 [56.8-92.2]%, p < .01). However, the median (IQR) of SARS-CoV-2 mixed peptides-specific T cell responses measured by ELISpot was significantly increased compared with baseline (30 [4-120] vs. 12 [0-56] T cells/106  PBMCs, p = .02) and not different from the controls. Our findings revealed weak HMI but comparable CMI responses in fully vaccinated KT recipients receiving inactivated SARS-CoV-2 vaccination compared to immunocompetent individuals (Thai Clinical Trials Registry, TCTR20210226002).
View publication
Establishment of a human iPSC line (SUTCMi001-A) derived from a healthy donor. L. Min et al. Stem cell research 2022 aug

Abstract

This study describes the characterization of one induced pluripotent stem cell line (iPSC) from a healthy female. It is crucial to use iPSCs derived from healthy individuals as controls in genetic disease studies. Thus, we established a human iPSC cell line derived from healthy people. The iPSC cell line was generated in our lab from the peripheral blood mononuclear cells (PBMCs) of a 28-year-old girl. The generated hiPSC line is free of episomal vectors, has a normal karyotype, expresses pluripotency markers and can differentiate into three germ layers in vivo.
View publication
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更多信息

更多信息
物种 人类
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • EasyEights™ EasySep™ Magnet (Catalog #18103) • Easy 50 EasySep™ Magnet (Catalog #18002)
样本来源 全血, 白细胞单采术样本, 白膜层, 脐带血, 骨髓
Selection Method Negative
标记抗体
  1. 抗人CD235（血型糖蛋白A/B）抗体，clone HIR2
    抗人CD235（血型糖蛋白A/B）抗体，clone HIR2

    小鼠Monoclonal IgG2b抗体，抗人糖蛋白A/B

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