Thioglycolate-elicited MΦs were isolated on day 4 from the peritoneum of mice injected intraperitoneally with 0.5 ml of 3% thioglycolate. Resident peritoneal MΦs were recovered from naive mice. BM-MΦs and BM-DCs were generated by culturing bone marrow cells with 20 ng ml−1 rM-CSF (PeproTech) for 7 days and with 16.67 ng ml−1 rGM-CSF (PeproTech) plus rIL-4 (5 ng ml−1, PeproTech) for 6 days, respectively. Cells were stimulated with TLR4-grade LPS from Escherichia Coli O111:B4 (Alexis Biochemicals). To enhance surface CD14 expression, BM-DCs were stimulated with 0.5 μM GpG (ODN 1826, Alexis Biochemicals) for 18 h. Splenic DCs were isolated by CD11c MACs beads (Miltenyi Biotec) and Marilyn cells were purified with CD4+ T cells isolation kit II (Miltenyi Biotec).
Cd4 t cells isolation kit 2
Manufactured by Miltenyi Biotec
The CD4+ T cells isolation kit II is a laboratory product designed to isolate CD4+ T cells from biological samples. The kit utilizes magnetic bead-based separation technology to specifically target and separate CD4+ T cells from other cell types present in the sample. This product allows for the efficient and reliable isolation of CD4+ T cells for downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
Ril 4, by Thermo Fisher Scientific (1 mentions)
Rgm csf, by Thermo Fisher Scientific (1 mentions)
Cd11c macs beads, by Miltenyi Biotec (1 mentions)
Rm csf, by Thermo Fisher Scientific (1 mentions)
Blood dendritic cell isolation kit 2, by Miltenyi Biotec (1 mentions)
Automacs cell separator, by Miltenyi Biotec (1 mentions)
Cd19 microbeads, by Miltenyi Biotec (1 mentions)
Nk cell isolation kit, by Miltenyi Biotec (1 mentions)
Cd14 microbead, by Miltenyi Biotec (1 mentions)
I0634, by Merck Group (1 mentions)
Tlrl smlps, by InvivoGen (1 mentions)
P1585 1mg, by Merck Group (1 mentions)
Tlrl fsl, by InvivoGen (1 mentions)
Tlrl pms, by InvivoGen (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Fugene, by Promega (1 mentions)
3 protocols using cd4 t cells isolation kit 2
1
Isolation and Culture of Murine Immune Cells
2
Isolation of Immune Cell Subsets
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CD4+ and CD8+ T-cells, NK-, B- and dendritic cells and monocytes were enriched from 50–100 x 106 PBMCs. Freshly isolated PBMCs were incubated with immunomagnetic beads—according to the manufacturer’s instructions—using CD4+ T Cells Isolation Kit II, CD8+ T Cells Isolation Kit II, CD19 MicroBeads, NK Cell Isolation Kit, Blood Dendritic Cell Isolation Kit II and CD14 MicroBeads (all from Miltenyi Biotec). Labeled PBMCs were applied to an AutoMACS cell-separator (Miltenyi Biotec) to isolate 2 x105 to 1 x 106 target cells.
3
Preparation and Activation of BMDMs, DCs, and CD4+ T Cells
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BMDMs and DCs were prepared as previously described (Vande Walle et al., 2014 (link)). The following reagents were used as indicated: FSL-1 (100 ng/ml; tlrl-fsl; InvivoGen), Pam3CSK4 (100 ng/ml; tlrl-pms; InvivoGen), S. minnesota LPS (100 ng/ml or 500 ng/ml; tlrl-smlps; InvivoGen), nigericin (20 µM, N7143-10MG; Sigma-Aldrich), C. difficile toxin B (1 µg/ml; #155L; List Biological Laboratories); Lipofectamine-2000 (5 µl; 11668–019; Invitrogen), Fugene (E2311; Promega). CD4+ T cells were purified by negative selection with CD4+ T cells isolation kit II (130–095-248; Miltenyi Biotec) according to manufacturer’s instructions. CD4+ T cell were activated with plate-bound anti-CD3 (4 µg/ml; 14–0031-85; eBioscience) and anti-CD28 antibodies (1 µg/ml; 14–0281-82; eBioscience), or with PMA (100 ng/ml; P1585-1MG; Sigma-Aldrich) together with ionomycin (1 µg/ml; I0634; Sigma-Aldrich).
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