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Dynabeads flowcomp kit

Manufactured by Thermo Fisher Scientific
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Dynabeads FlowComp Kit is a magnetic bead-based product designed for the isolation and enrichment of specific cell populations from complex biological samples. The kit utilizes magnetic beads coated with antibodies or ligands that bind to target cells, allowing for their separation and purification using a magnetic field.

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Lab products found in correlation

Mil 4, by Thermo Fisher Scientific (2 mentions) Dynabeads mouse t activator cd3 cd28, by Thermo Fisher Scientific (2 mentions) Dynabeads mouse cd43, by Thermo Fisher Scientific (2 mentions) Gm csf, by Cytek Biosciences (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Cd11c microbeads, by Miltenyi Biotec (2 mentions)

3 protocols using dynabeads flowcomp kit

1

Isolation and Activation of Immune Cells

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Primary dendritic cells, B cells, CD4+ T cells, and macrophages from C57BL/6J wild-type and miR-155 KO mice were cultured in RPMI medium with 10% FBS. Prior to harvesting primary dendritic cells, mice were subcutaneously injected with 1 × 106 B16 melanoma cells expressing Flt3 ligand for about two weeks. After purification of splenic CD11c+ dendritic cells by CD11c microbeads (Miltenyi Biotec), dendritic cells were activated in a medium containing 100 ng/ml LPS (SIGMA) and 20 ng/ml GM-CSF (Tonbo). Splenic primary B cells were purified by negative selection using Dynabeads Mouse CD43 (Invitrogen), and activated in a medium containing 25 μg/ml LPS and 6.5 ng/ml mIL-4 (PeproTech). CD4+ T cells from lymph node and spleen were purified with Dynabeads FlowComp Kit (Invitrogen). CD4+CD25−CD44− T cells were then activated with Dynabeads Mouse T-Activator CD3/CD28 (Invitrogen). Intraperitoneal macrophages, induced by thioglycollate injection, were harvested and activated with 100 ng/ml LPS.
Hsin J.P., Lu Y., Loeb G.B., Leslie C.S, & Rudensky A.Y. (2018). The effect of cellular context on miR-155-mediated gene regulation in four major immune cell types. Nature immunology, 19(10), 1137-1145.
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2

Isolation and Activation of Immune Cells

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Primary dendritic cells, B cells, CD4+ T cells, and macrophages from C57BL/6J wild-type and miR-155 KO mice were cultured in RPMI medium with 10% FBS. Prior to harvesting primary dendritic cells, mice were subcutaneously injected with 1 × 106 B16 melanoma cells expressing Flt3 ligand for about two weeks. After purification of splenic CD11c+ dendritic cells by CD11c microbeads (Miltenyi Biotec), dendritic cells were activated in a medium containing 100 ng/ml LPS (SIGMA) and 20 ng/ml GM-CSF (Tonbo). Splenic primary B cells were purified by negative selection using Dynabeads Mouse CD43 (Invitrogen), and activated in a medium containing 25 μg/ml LPS and 6.5 ng/ml mIL-4 (PeproTech). CD4+ T cells from lymph node and spleen were purified with Dynabeads FlowComp Kit (Invitrogen). CD4+CD25−CD44− T cells were then activated with Dynabeads Mouse T-Activator CD3/CD28 (Invitrogen). Intraperitoneal macrophages, induced by thioglycollate injection, were harvested and activated with 100 ng/ml LPS.
Hsin J.P., Lu Y., Loeb G.B., Leslie C.S, & Rudensky A.Y. (2018). The effect of cellular context on miR-155-mediated gene regulation in four major immune cell types. Nature immunology, 19(10), 1137-1145.
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3

Isolation and Analysis of IL10-Deficient Tregs

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To test the deletion of IL10 in T reg cells, we prepared single cell suspensions from spleens of Il10flox/flox and IL10flox/floxFoxp3YFP-Cre x mice and isolated CD4+CD25+ Treg cells using the Dynabeads FlowComp kit (Invitrogen Life Technologies). This method uses negative selection of CD4+ T cells, followed by positive selection of CD25+ regulatory T cells. First, an antibody mixture was added to the spleen single cell suspension to bind the non CD4+ T cells. Mouse Depletion Dynabeads were then added and the bound non CD4+ cells were removed with a magnet. The remaining untouched mouse CD4+ T-cells were then labeled with CD25 antibody and positively isolated using Dynabeads. The T reg cells adherent to Dynabeads were separated with a magnet and used for qRT-PCR to measure the IL10 transcript levels.
Wang W.W., Wang Y., Li K., Tadagavadi R., Friedrichs W.E., Budatha M, & Reeves W.B. (2020). IL-10 from dendritic cells but not from T regulatory cells protects against cisplatin-induced nephrotoxicity. PLoS ONE, 15(9), e0238816.
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