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Complete Kit for Human Whole Blood CD34+ Cells

Immunomagnetic positive selection kit

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From: 747 USD

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Immunomagnetic positive selection kit
From: 747 USD

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Overview

The Complete Kit for Human Whole Blood CD34+ Cells is a simple two-step method that is designed to isolate CD34+ cells from whole blood or buffy coat.

First, hematopoietic progenitor cells are pre-enriched using the RosetteSep™ Human Hematopoietic Progenitor Cell Enrichment Cocktail (15186C) with antibodies recognizing CD2, CD3, CD14, CD16, CD19, CD24, CD56, CD66b and CD61 surface markers. CD34+ cells are then selected using the EasySep™ Human CD34 Positive Selection Cocktail (18066C), which contains an antibody recognizing CD34. RosetteSep™ binds unwanted cells to red blood cells (RBCs), forming immunorosettes, which sediment during density gradient centrifugation. The pre-enriched fraction containing CD34+ cells is harvested from the interface between the plasma and density gradient medium. The pre-enriched CD34+ cells are then labeled with antibodies and magnetic particles, and separated without columns using an EasySep™ magnet. Unwanted cells are simply poured off, while desired cells remain in the tube. Isolated CD34+ cells are immediately available for downstream applications.
Advantages:
• Fast and easy-to-use
• Up to 98% purity
• No columns required
• Can be combined with SepMate™ for consistent, high-throughput sample processing
Components:
  • Complete Kit for Human Whole Blood CD34+ Cells (Catalog #15086)
    • RosetteSep™ Human Hematopoietic Progenitor Cell Enrichment Cocktail, 3 x 2 mL
    • EasySep™ Human CD34 Positive Selection Cocktail, 0.4 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
  • RoboSep™ Complete Kit for Human WB CD34 (Catalog #15086RF)
    • RosetteSep™ Human Hematopoietic Progenitor Cell Enrichment Cocktail, 3 x 2 mL
    • EasySep™ Human CD34 Positive Selection Cocktail, 0.4 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
Magnet Compatibility:
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• RoboSep™-S (Catalog #21000)
Subtype:
Cell Isolation Kits
Cell Type:
Hematopoietic Stem and Progenitor Cells
Species:
Human
Sample Source:
Buffy Coat; Whole Blood
Selection Method:
Positive
Application:
Cell Isolation
Brand:
EasySep; RoboSep; RosetteSep
Area of Interest:
Immunology; Stem Cell Biology

Scientific Resources

Educational Materials

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Frequently Asked Questions

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.
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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.

Data and Publications

Data

Figure 1.Typical CD34 Selection Profile on Human Whole Blood

Starting with whole peripheral blood, the CD34+ cell content of the isolated fraction is typically 95.1 ± 4.5% (gated on viable CD45+ cells; mean ± SD for the silver “The Big Easy” EasySep™ Magnet). In the above example, the purities of the start and final isolated fractions are 0.06% and 96.2%, respectively.

Publications

(1)
Cell reports 2016 NOV

Nucleosome Density ChIP-Seq Identifies Distinct Chromatin Modification Signatures Associated with MNase Accessibility.

Lorzadeh A et al.

Abstract

Nucleosome position, density, and post-translational modification are widely accepted components of mechanisms regulating DNA transcription but still incompletely understood. We present a modified native ChIP-seq method combined with an analytical framework that allows MNase accessibility to be integrated with histone modification profiles. Application of this methodology to the primitive (CD34+) subset of normal human cord blood cells enabled genomic regions enriched in one versus two nucleosomes marked by histone 3 lysine 4 trimethylation (H3K4me3) and/or histone 3 lysine 27 trimethylation (H3K27me3) to be associated with their transcriptional and DNA methylation states. From this analysis, we defined four classes of promoter-specific profiles and demonstrated that a majority of bivalent marked promoters are heterogeneously marked at a single-cell level in this primitive cell type. Interestingly, extension of this approach to human embryonic stem cells revealed an altered relationship between chromatin modification state and nucleosome content at promoters, suggesting developmental stage-specific organization of histone methylation states.
STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.