When isolating cells with Dynabeads magnetic beads, what is more important: bead-to-target cell ratio or the concentration of beads in the bead/cell mixture?
Both bead-to-target cell ratio and the concentration of beads in the bead/cell mixture are important and should be considered. For example, when using the Dynabeads magnetic beads M-450 CD4 positive isolation or depletion kit, a 4:1 bead-to-target cell ratio should be maintained. To capture 95% of target cells for molecular applications, the bead concentration must always be 1 x 10e7 beads per milliliter of sample. To deplete 99% CD4 cells from the starting sample, the bead concentration must always be 2 x 10e7 beads per milliliter of sample. Please consult the package insert for recommended bead concentrations of each product.
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Can I perform flow cytometry analysis on cells labeled with Dynabeads beads?
No, the data are not ideal when cells have beads attached to them. You need to use one of the Dynabeads Assays that allow you to remove the beads from your cells for downstream analysis. These include the Dynabeads FlowComp assays and DETACHaBEAD assays.
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Which Dynabeads kits will yield bead-free cells that can be used for flow cytometry?
Bead-free cells can be isolated using Dynabeads kits that include DETACHaBEAD technology. These include the Dynabeads FlowComp kits and Dynabeads CELLection Cell Isolation Kits. Also, any of the negative isolation kits will generate cells that are bead-free.
Human Cells
Dynabeads FlowComp Human CD4 Kit (T cells) (Cat. No. 11361D)
Dynabeads FlowComp Human CD8 Kit (T cells) (Cat. No. 11362D)Dynabeads FlowComp Human CD3 Kit (T cells) (Cat. No. 11365D)
Dynabeads FlowComp Human CD14 Kit (monocytes) (Cat. No. 11367D)
Dynabeads CD34 Positive Isolation Kit (hematopoietic progenitor cells) (Cat. No. 11301D)
Dynabeads CD4 Positive Isolation Kit (T cells) (Cat. No. 11331D)
Dynabeads CD8 Positive Isolation Kit (T cells) (Cat. No. 11333D)
Dynabeads Regulatory CD4+/CD25+ T Cell Kit (Cat. No. 11363D)
DETACHaBEAD CD19 Kit (Cat. No. 12506D)
CELLection Epithelial Enrich Dynabeads (Cat. No. 16203)
Dynabeads Human Dentritic Cells (DC) Enrichment Kit (Cat. No. 11308D)
Mouse Cells
Dynabeads FlowComp Mouse CD4 Kit (T cells) (Cat. No. 11461D)
Dynabeads FlowComp Mouse CD8 Kit (T cells) (Cat. No. 11462D)
Dynabeads FlowComp Mouse CD4+CD25+ Treg Cells Kit (Cat. No. 11463D)
Dynabeads FlowComp Mouse Pan T (CD90.2) Kit (Cat. No. 11465D)
Dynabeads Mouse Pan T (Thy1.2) (Cat. No. 11443D)
DETACHaBEAD Mouse CD4 Kit (T cells) (Cat. No. 12406D)
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How do I convert the Dynabeads magnetic bead concentration from 1 mg beads/mL to actual bead numbers per milliliter?
This depends on the bead size. In general, for M-450 Dynabeads, 1 mg/mL beads is approximately equivalent to 1.3 X 10E7 beads/mL; for M-280 Dynabeads, 1 mg/mL beads is approximately equivalent to 6.7 X 10E7 beads/mL.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Will Dynabeads magnetic beads be internalized if cultured with the beads on?
Whether cells will internalize the Dynabeads magnetic beads during culture will depend on the cell type. Due to the bead size (usually 4.5µm in diameter) Dynabeads magnetic beads will not be internalized into the endocytic pathway e.g., via clathrin coated pits. The clathrin coated pits are typically not more than 500 nm in size, which is far too small for endocytosis of the beads. However, if cells with phagocytic activities (e.g., monocytes/macrophages) are present, the Dynabeads magnetic beads will be phagocytosed into the phagolysosomes by these specialized cells. So it would really depend on the cell type.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
