Peripheral blood mononuclear cells (MNCs) were separated by gradient centrifugation (800g, 30 min) using Ficoll (Ficoll-Paque; GE Healthcare Biosciences). MNCs were collected and freed from plasma by re-suspending the cells in PBS with 2% FBS and 1 mM EDTA, and centrifugation. MNCs were either used for flow cytometry analysis of lineage-negative (Lin−) or Lin−CD45lowCD34+ HSPC populations, or for the enrichment of CD34+ HSPCs. A negative selection kit (StemCell Technologies) was used for the enrichment of Lin− cells by following supplier’s instructions. This kit depletes cells that are positive for CD3, CD14, CD16, CD19, CD20, and CD56, by using magnetic microparticles. Lin− cells were further enriched for CD34+ cells by positive selection by using an immunomagnetic separation kit (StemCell Technologies) as per manufacturer’s instructions. Cells were either used for experiments involving hypoxia or preserved at − 80 °C for enzyme activity assays.
Immunomagnetic separation kit
Manufactured by STEMCELL
The Immunomagnetic Separation Kit is a tool used to isolate specific cell types from complex samples. It utilizes magnetic beads coated with antibodies to selectively bind and separate target cells from a heterogeneous mixture. The kit provides a simple and efficient method for cell isolation, enabling researchers to enrich and purify cell populations for downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using immunomagnetic separation kit
1
Isolation and Enrichment of HSPC
2
Isolation and Enrichment of Human HSPCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peripheral blood mononuclear cells (MNCs) were separated by gradient centrifugation (800×g , 30 min) using Ficoll (Ficoll-Paque; GE Healthcare Biosciences). MNCs were either used for flow cytometry analysis of lineage-negative (Lin−) or Lin−CD45lowCD34+ HSPC populations, or for the enrichment of CD34+ HSPCs. A negative selection kit (Stemcell Technologies) was used for the enrichment of Lin− cells by following supplier’s instructions, which involves depletion of cells expressing CD3, CD14, CD16, CD19, CD20, and CD56. Lin− cells were further enriched for CD34+ cells by positive-selection by using immunomagnetic separation kit (Stemcell Technologies) as per manufacturer’s instructions. Cells were either used for experiments in vitro or processed for in vivo testing in mice.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!