EasySep™ Human Cord Blood CD34 Positive Selection Kit II
Immunomagnetic positive selection kit
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EasySep™ Human Cord Blood CD34 Positive Selection Kit II -
RoboSep™ Human CB CD34 Positive Selection Kit II -
Label for EasySep™ Human Cord Blood CD34 Positive Selection Kit II
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Required Products
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Density gradient medium for the isolation of mononuclear cells -
Magnet for column-free immunomagnetic separation -
Cell separation buffer -
Compatible antibodies for purity assessment of isolated cells
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).
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
• 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
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EasySep™ Human Cord Blood CD34 Positive Selection Kit III
Immunomagnetic positive selection kit -
Tube for density gradient centrifugation for in vitro diagnostic (IVD) applications
Labeling Antibodies
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Anti-Human CD34 Antibody, Clone 8G12
Mouse monoclonal IgG1 antibody against human CD34 -
Anti-Human CD45 Antibody, Clone HI30
Mouse monoclonal IgG1 antibody against human, chimpanzee CD45 -
Anti-Human CD34 Antibody, Clone 581
Mouse monoclonal IgG1 antibody against human CD34
Scientific Resources
Product Documentation
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(8)
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.
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.
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Data and Publications
Data
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.
Abstract
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.
Abstract
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.
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.