EasySep™ Human Cord Blood CD34 Positive Selection Kit II

Immunomagnetic positive selection kit
Catalog #
17896_C
Immunomagnetic positive selection kit
From: 656 USD
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Required Products
  1. Lymphoprep™|07851
    Lymphoprep™

    Density gradient medium for the isolation of mononuclear cells

  2. EasySep™ Magnet
    EasySep™ Magnet

    Magnet for column-free immunomagnetic separation

  3. EasySep™ Buffer
    EasySep™ Buffer

    Cell separation buffer

Overview

Isolate highly purified CD34+ cells from fresh whole umbilical cord blood using a simple, two-step procedure. First, hematopoietic progenitor cells are pre-enriched using the RosetteSep™ Human Cord Blood CD34 Pre-Enrichment Cocktail (15896C) with antibodies recognizing T cell, B cell, myeloid cell and platelet surface markers. CD34+ cells are then selected using the EasySep™ Human CD34 Positive Selection Cocktail (18096C), which contains an antibody recognizing CD34.

If isolating CD34+ cells from fresh blood or buffy coat the Complete Kit for Human Whole Blood CD34+ Cells (Catalog #15086) is recommended.

If isolating CD34+ cells from other samples, including fresh or previously frozen mobilized peripheral blood or bone marrow mononuclear cells, or from previously frozen cord blood mononuclear cells, we recommend using the EasySep™ Human CD34 Positive Selection Kit (Catalog #17856).

This product replaces the stand alone EasySep™ Human Cord Blood CD34 Positive Selection Kit (Catalog #18096).
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
  • EasySep™ Human Cord Blood CD34 Positive Selection Kit II (Catalog #17896)
    • RosetteSep™ Cord Blood CD34 Pre-Enrichment Cocktail II, 2 x 2.5 mL
    • EasySep™ Human CD34 Positive Selection Cocktail, 2 x 1 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
  • RoboSep™ Human Cord Blood CD34 Positive Selection Kit II (Catalog #17896RF)
    • RosetteSep™ Cord Blood CD34 Pre-Enrichment Cocktail II, 2 x 2.5 mL
    • EasySep™ Human CD34 Positive Selection Cocktail, 2 x 1 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125) x 2
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• RoboSep™-S (Catalog #21000)
Subtype
Cell Isolation Kits
Cell Type
Hematopoietic Stem and Progenitor Cells
Species
Human
Sample Source
Cord Blood
Selection Method
Positive
Application
Cell Isolation
Brand
EasySep, RoboSep, RosetteSep
Area of Interest
Immunology, Stem Cell Biology

Related Products

Labeling Antibodies

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
EasySep™ Human Cord Blood CD34 Positive Selection Kit II
Catalog #
17896
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ Human CB CD34 Positive Selection Kit II
Catalog #
17896RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human Cord Blood CD34 Positive Selection Kit II
Catalog #
17896
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human Cord Blood CD34 Positive Selection Kit II
Catalog #
17896
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Human Cord Blood CD34 Positive Selection Kit II
Catalog #
17896
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human CB CD34 Positive Selection Kit II
Catalog #
17896RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human CB CD34 Positive Selection Kit II
Catalog #
17896RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human CB CD34 Positive Selection Kit II
Catalog #
17896RF
Lot #
All
Language
English

Educational Materials(8)

Brochure
Hematopoietic Stem and Progenitor Cells - Products for Your Research
Brochure
EasySep™ Cell Separation Technology
Technical Bulletin
Isolation of CD34+ Cells from Human Cord Blood
Wallchart
Human Immune Cytokines
Video
1:13
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
Video
0:57
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
Video
1:37
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
Webinar
45:17
Troubleshooting In Vitro Expansion of Leukemic Cells

Frequently Asked Question

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.
Read More

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

EasySep™ Human Cord Blood CD34 Positive Selection Kit II

Figure 1. EasySep™ Human Cord Blood CD34 Positive Selection Kit II

Starting with fresh cord blood, the CD34+ cell content of the isolated fraction is typically 91 ± 9% (mean ± SD using the purple EasySep™ Magnet).

Publications (2)

Clinical cancer research : an official journal of the American Association for Cancer Research 2020 feb Discovery of Berberine that Targetedly Induces Autophagic Degradation of both BCR-ABL and BCR-ABL T315I through Recruiting LRSAM1 for Overcoming Imatinib Resistance. Z. Yin et al.

Abstract

PURPOSE Imatinib, the breakpoint cluster region protein (BCR)/Abelson murine leukemia viral oncogene homolog (ABL) inhibitor, is widely used to treat chronic myeloid leukemia (CML). However, imatinib resistance develops in many patients. Therefore, new drugs with improved therapeutic effects are urgently needed. Berberine (BBR) is a potent BCR-ABL inhibitor for imatinib-sensitive and -resistant CML. EXPERIMENTAL DESIGN Protein structure analysis and virtual screening were used to identify BBR targets in CML. Molecular docking analysis, surface plasmon resonance imaging, nuclear magnetic resonance assays, and thermoshift assays were performed to confirm the BBR target. The change in BCR-ABL protein expression after BBR treatment was assessed by Western blotting. The effects of BBR were assessed in vitro in cell lines, in vivo in mice, and in human CML bone marrow cells as a potential strategy to overcome imatinib resistance. RESULTS We discovered that BBR bound to the protein tyrosine kinase domain of BCR-ABL. BBR inhibited the activity of BCR-ABL and BCR-ABL with the T315I mutation, and it also degraded these proteins via the autophagic lysosome pathway by recruiting E3 ubiquitin-protein ligase LRSAM1. BBR inhibited the cell viability and colony formation of CML cells and prolonged survival in CML mouse models with imatinib sensitivity and resistance. CONCLUSIONS The results show that BBR directly binds to and degrades BCR-ABL and BCR-ABL T315I via the autophagic lysosome pathway by recruiting LRSAM1. The use of BBR is a new strategy to improve the treatment of patients with CML with imatinib sensitivity or resistance.
Nature Biotechnology 2016 APR Directed evolution of a recombinase that excises the provirus of most HIV-1 primary isolates with high specificity. Karpinski J et al.

Abstract

Current combination antiretroviral therapies (cART) efficiently suppress HIV-1 reproduction in humans, but the virus persists as integrated proviral reservoirs in small numbers of cells. To generate an antiviral agent capable of eradicating the provirus from infected cells, we employed 145 cycles of substrate-linked directed evolution to evolve a recombinase (Brec1) that site-specifically recognizes a 34-bp sequence present in the long terminal repeats (LTRs) of the majority of the clinically relevant HIV-1 strains and subtypes. Brec1 efficiently, precisely and safely removes the integrated provirus from infected cells and is efficacious on clinical HIV-1 isolates in vitro and in vivo, including in mice humanized with patient-derived cells. Our data suggest that Brec1 has potential for clinical application as a curative HIV-1 therapy.
View All Publications

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