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RosetteSep™HLA淋巴细胞富集试剂盒

免疫密度阴性选择鸡尾酒
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产品号 #（选择产品）

产品号 #15271HLA_C

免疫密度阴性选择鸡尾酒

产品优势

  • 快速且易于使用
  • 不需要特殊设备或培训
  • 分离的细胞不受影响

产品组分包括

  • RosetteSep™HLA淋巴细胞富集试剂盒（目录#15271HLA）
    • RosetteSep™HLA淋巴细胞浓缩鸡尾酒，5 × 2ml
    • RosetteSep™DM-L,（目录#15705）x 2
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Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
  1. Lymphoprep™
    Lymphoprep™

    Density gradient medium for the isolation of mononuclear cells

  2. Dulbecco's Phosphate Buffered Saline with 2% Fetal Bovine Serum
    Dulbecco's Phosphate Buffered Saline with 2% ...

    Cell culture 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

概述

RosetteSep™HLA淋巴细胞富集鸡尾酒旨在富集淋巴细胞（CD3）+)全血细胞通过负选择进行谱系特异性嵌合分析。四聚体抗体复合物识别红细胞（红细胞）上的CD16、CD19、CD20、CD36、CD56和糖蛋白A，以清除不需要的细胞为目标。当在浮性密度介质上离心时，如RosetteSep™DM-L（目录#15705），不需要的细胞颗粒与红细胞一起。纯化后的淋巴样细胞在等离子体和浮力密度介质之间的界面上以高度富集的群体存在。所需的细胞立即准备好进行嵌合分析或其他下游应用。请注意：根据欧洲议会和理事会关于体外诊断医疗器械的新欧盟法规2017/746 (IVDR)，该法规预计将于2022年生效，STEMCELL最近注销并删除了与这些产品相关的CE IVD声明，并且它们现在标记为“仅供研究使用”。请注意，与这些产品有关的形式、功能和质量保证没有任何变化。如欲了解更多信息，请与质量保证及监管部联系qaschangenote@stemcell.com.

Subtype
Cell Isolation Kits
 
Cell Type
T Cells
 
Species
Human
 
Sample Source
Buffy Coat, Whole Blood
 
Selection Method
Negative
 
Application
Cell Isolation, In Vitro Diagnostic
 
Brand
RosetteSep
 
Area of Interest
Chimerism, HLA, Immunology
 

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Data Figures

FACS Histogram Results with RosetteSep™ HLA Lymphoid Cell Enrichment Kit

Figure 1. FACS Histogram Results with RosetteSep™ HLA Lymphoid Cell Enrichment Kit

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet 1
Product Name
RosetteSep™ HLA Lymphoid Cell Enrichment Kit
Catalog #
15271HLA
Lot #
Lots above 1000075249
Language
English
Document Type
Product Information Sheet 2
Product Name
RosetteSep™ HLA Lymphoid Cell Enrichment Kit
Catalog #
15271HLA
Lot #
Lots above 1000075249
Language
Multi
Document Type
Safety Data Sheet
Product Name
RosetteSep™ HLA Lymphoid Cell Enrichment Kit
Catalog #
15271HLA
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (4)

Cell Separation Products for HLA Analysis
Brochure
Cell Separation Products for HLA Analysis
Human Immune Cytokines
Wallchart
Human Immune Cytokines
Isolate Highly Purified Cells for HLA Analysis with RosetteSep™ Immunodensity Cell Isolation
1:21
Video
Isolate Highly Purified Cells for HLA Analysis with RosetteSep™ Immunodensity Cell Isolation
Isolate Cells from Whole Blood without Columns or Magnets: RosetteSep™ Immunodensity Cell Separation
2:19
Video
Isolate Cells from Whole Blood without Columns or Magnets: RosetteSep™ Immunodensity Cell Separati...

Frequently Asked Questions

What is RosetteSep™?

RosetteSep™ is a rapid cell separation procedure for the isolation of purified cells directly from whole blood, without columns or magnets.

How does RosetteSep™ work?

The antibody cocktail crosslinks unwanted cells to red blood cells (RBCs), forming rosettes. The unwanted cells then pellet with the free RBCs when centrifuged over a density centrifugation medium (e.g. Ficoll-Paque™ PLUS, Lymphoprep™).

What factors affect cell recovery?

The temperature of the reagents can affect cell recovery. All reagents should be at room temperature (sample, density centrifugation medium, PBS, centrifuge) before performing the isolations. Layering can also affect recovery so be sure to carefully layer the sample to avoid mixing with the density centrifugation medium as much as possible. Be sure to collect the entire enriched culture without disturbing the RBC pellet. A small amount of density centrifugation medium can be collected without worry.

Which cell samples can RosetteSep™ be used with?

RosetteSep™ can be used with leukapheresis samples, bone marrow or buffy coat, as long as: the concentration of cells does not exceed 5 x 107 per mL (can dilute if necessary); and there are at least 100 RBCs for every nucleated cell (RBCs can be added if necessary).

Can RosetteSep™ be used with previously frozen or cultured cells?

Yes. Cells should be re-suspended at 2 - 5 x 107 cells / mL in PBS + 2% FBS. Fresh whole blood should be added at 250 µL per mL of sample, as a source of red cells.

Can RosetteSep™ be used to enrich progenitors from cord blood?

Yes. Sometimes cord blood contains immature nucleated red cells that have a lower density than mature RBCs. These immature red cells do not pellet over Ficoll™, which can lead to a higher RBC contamination than peripheral blood separations.

Does RosetteSep™ work with mouse cells?

No, but we have developed EasySep™, a magnetic-based cell isolation system which works with mouse and other non-human species.

Which anticoagulant should be used with RosetteSep™?

Peripheral blood should be collected in heparinized Vacutainers. Cord blood should be collected in ACD.

Should the anticoagulant be washed off before using RosetteSep™?

No, the antibody cocktail can be added directly to the sample.

Publications (2)

Dendritic cells induce MUC1 expression and polarization on human T cells by an IL-7-dependent mechanism. Vasir B et al. Journal of immunology (Baltimore, Md. : 1950) 2005 FEB

Abstract

The MUC1 transmembrane mucin is expressed on the surface of activated human T cells; however, the physiologic signals responsible for the regulation of MUC1 in T cells are not known. The present studies demonstrate that IL-7, but not IL-2 or IL-4, markedly induces MUC1 expression on CD3+ T cells. MUC1 was also up-regulated by IL-15, but to a lesser extent than that found with IL-7. The results show that IL-7 up-regulates MUC1 on CD4+, CD8+, CD25+, CD69+, naive CD45RA+, and memory CD45RO+ T cells. In concert with induction of MUC1 expression by IL-7, activated dendritic cells (DC) that produce IL-7 up-regulate MUC1 on allogeneic CD3+ T cells. DC also induce MUC1 expression on autologous CD3+ T cells in the presence of recall Ag. Moreover, DC-induced MUC1 expression on T cells is blocked by a neutralizing anti-IL-7 Ab. The results also demonstrate that DC induce polarization of MUC1 on T cells at sites opposing the DC-T cell synapse. These findings indicate that DC-mediated activation of Ag-specific T cells is associated with induction and polarization of MUC1 expression by an IL-7-dependent mechanism.
View publication
Rapid induction of complete donor chimerism by the use of a reduced-intensity conditioning regimen composed of fludarabine and melphalan in allogeneic stem cell transplantation for metastatic solid tumors. Ueno NT et al. Blood 2003 NOV

Abstract

We evaluated the feasibility and efficacy of a reduced-intensity conditioning (RIC) regimen of fludarabine and melphalan to achieve rapid complete donor chimerism after allogeneic stem cell transplantation (SCT) in patients with metastatic solid tumors. Between January 1999 and January 2003, 8 patients with metastatic breast cancer (BC) and 15 with metastatic renal cell carcinoma (RCC) underwent allogeneic SCT after an RIC regimen of 5 days of fludarabine and 2 days of melphalan. Filgrastim-mobilized stem cells from HLA-identical related or unrelated donors were infused. Prophylaxis for graft-versus-host disease (GVHD) consisted of tacrolimus and methotrexate. All 22 evaluable patients had 100% donor chimerism at day 30 and at all measurement times thereafter. One patient died 19 days after SCT. Nine patients (39%) had grades II to IV acute GVHD and 10 patients (43%) had chronic GVHD. Five patients (22%) died of nonrelapse treatment-related complications. Treatment-related disease response was seen in 10 patients (45%), with 3 complete responses, 2 partial responses, and 5 minor responses. Fludarabine-melphalan is a feasible and effective RIC regimen for allogeneic SCT in metastatic BC and RCC. It induces rapid complete donor chimerism without the need for donor lymphocyte infusion. Tumor regression associated with GVHD is consistent with graft-versus-tumor effect.
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更多信息

更多信息
Species Human
Sample Source Buffy Coat, Whole Blood
Selection Method Negative
标记抗体
  1. 抗人CD3抗体克隆UCHT1
    抗人CD3抗体克隆UCHT1

    小鼠（BALB/c）抗人、黑猩猩CD3单克隆IgG1抗体

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