EasySep™ HLA Chimerism Whole Blood CD3 Positive Selection Kit

Immunomagnetic positive selection cell isolation kit

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Immunomagnetic positive selection cell isolation kit
From: 659 USD

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Overview Related Products Scientific Resources Product Applications Data and Publications

Overview

The EasySep™ HLA Chimerism Whole Blood CD3 Positive Selection Kit is designed to isolate CD3+ cells from fresh whole blood or buffy coat by positive selection. Desired cells are targeted with Tetrameric Antibody Complexes recognizing CD3 and dextran-coated magnetic particles. The cocktail also contains an antibody to human Fc receptor to minimize nonspecific binding. Labeled cells are separated using an EasySep™ magnet without the use of columns. Cells of interest remain in the tube while unwanted cells are poured off. The CD3 antigen is expressed on T cells and NKT cells.

This product replaces the EasySep™ Human Whole Blood CD3 Positive Selection Kit (Catalog #18081) and EasySep™ HLA Whole Blood CD3 Positive Selection Kit (Catalog #18081HLA), for even faster cell isolations.
Advantages:
• Fast and easy-to-use
• Up to 99% purity
• No columns required
• Compatible across EasySep™ "The Big Easy", EasyEights™, and RoboSep™-S platforms
Components:
  • EasySep™ HLA Chimerism Whole Blood CD3 Positive Selection Kit (Catalog #17871)
    • EasySep™ HLA Chimerism Whole Blood CD3 Positive Selection Cocktail, 3 x 1 mL
    • EasySep™ Dextran RapidSpheres™ 50101, 3 x 1 mL
    • EasySep™ RBC Lysis Buffer 10X Concentrate, 10 mL (Catalog #20110)
  • RoboSep™ HLA Chimerism Whole Blood CD3 Positive Selection Kit with Filter Tips (Catalog #17871RF)
    • EasySep™ HLA Chimerism Whole Blood CD3 Positive Selection Cocktail, 3 x 1 mL
    • EasySep™ Dextran RapidSpheres™ 50101, 3 x 1 mL
    • EasySep™ RBC Lysis Buffer 10X Concentrate, 10 mL (Catalog #20110)
    • RoboSep™ Buffer (Catalog #20104) x 2
RoboSep™ Filter Tips (Catalog #20125) x 2
Magnet Compatibility:
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• RoboSep™-S (Catalog #21000)
 
Subtype:
Cell Isolation Kits
Cell Type:
T Cells
Species:
Human
Sample Source:
Buffy Coat; Whole Blood
Selection Method:
Positive
Brand:
EasySep
Area of Interest:
HLA; Immunology

Scientific Resources

Product Documentation

Educational Materials

(12)
Brochure
EasySep™ Cell Separation Technology
Brochure
HLA Antibodies for Purity Assessment
Brochure
Cell Separation Products for HLA Analysis
Technical Bulletin
Purity Assessment by Flow Cytometry for Lineage-Specific Chimerism Analysis
Technical Bulletin
Fully Automated, High Purity Cell Isolation for Chimerism Labs
Wallchart
Frequencies of Cell Types in Human Peripheral Blood
Wallchart
Human Immune Cytokines
Video
4:55
How to Recover Cells from a Leukocyte Reduction System (LRS) Cone
Video
1:41
Automate Cell Isolation with the RoboSep™-S Cell Separation Instrument
Video
1:57
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Webinar
56:02
Fully Automated Cell Separation for Chimerism Analysis with RoboSep™
Scientific Poster
Isolation of Very Pure Lymphoid (CD3+, CD19+ or CD56+) and Myeloid (CD15+ or CD33/CD66b+) Cell Subsets in as Little as 15 Minutes for Use in Lineage-Specific Chimerism Analysis

Product 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 for
Workflow Stages
T Cell Research
Cell Sourcing
Cell Isolation
Cell Activation and Expansion
Cell Characterization
Chimerism Analysis
Cell Isolation
Characterization

Data and Publications

Data

Starting with human whole blood, the CD3+ cell content of the isolated fraction typically ranges from 92.4 - 99.8% (as assessed by staining the start and isolated fractions with anti-CD3 or anti-CD2 antibodies, respectively). In the above example, the purities of the start and the final isolated fractions are 18.2% and 98.1%, respectively (gated on CD45).
NOTE: Red blood cells were removed from the start sample by lysis prior to flow cytometry.

Publications

(1)
Journal of immunology (Baltimore, Md. : 1950) 2018 NOV

Neutrophil and Granulocytic Myeloid-Derived Suppressor Cell-Mediated T Cell Suppression Significantly Contributes to Immune Dysregulation in Common Variable Immunodeficiency Disorders.

M. Vlkova et al.
Abstract
View Publication

Abstract

Common variable immunodeficiency disorders (CVID) represent a group of primary immunodeficiency diseases characterized by hypogammaglobulinemia and impaired specific Ab response, resulting in recurrent infections due to dysfunctional immune response. The specific mechanisms mediating immune deficiency in CVID remain to be determined. Previous studies indicated that immune dysregulation in CVID patients is associated with chronic microbial translocation, systemic immune activation, and altered homeostasis of lymphocytic and myeloid lineages. A detailed phenotypic, functional characterization of plasma markers and immune cell populations was performed in 46 CVID patients and 44 healthy donors. CVID patients displayed significantly elevated plasma levels of a marker of neutrophil activation neutrophil gelatinase-associated lipocalin. Neutrophils from CVID patients exhibited elevated surface levels of CD11b and PD-L1 and decreased levels of CD62L, CD16, and CD80, consistent with a phenotype of activated neutrophils with suppressive properties. Neutrophils from CVID patients actively suppressed T cell activation and release of IFN-$\gamma$ via the production of reactive oxygen species. Furthermore, CVID was associated with an increased frequency of low-density neutrophils (LDNs)/granulocytic myeloid-derived suppressor cells. LDN/granulocytic myeloid-derived suppressor cell frequency in CVID patients correlated with reduced T cell responsiveness. Exogenous stimulation of whole blood with bacterial LPS emulated some but not all of the phenotypic changes observed on neutrophils from CVID patients and induced neutrophil population with LDN phenotype. The presented data demonstrate that neutrophils in the blood of CVID patients acquire an activated phenotype and exert potent T cell suppressive activity. Specific targeting of myeloid cell-derived suppressor activity represents a novel potential therapeutic strategy for CVID.
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.