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.
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.
Data Figures
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
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.
Resources and Publications
Educational Materials (5)
Frequently Asked Questions
Publications (1)
Nucleosome Density ChIP-Seq Identifies Distinct Chromatin Modification Signatures Associated with MNase Accessibility. Cell reports 2016 NOV
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.
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.
