Spleens from BALB/c animals were disrupted mechanically. CD4+ T cells were obtained by negative selection using a cocktail of Abs containing the following culture supernatants (produced in house): B220 (RA3-6B2), F4/80 (HB-198), MHC-II (TIB 120), NK1.1 (HB-191) and CD8 (2.43). After supernatant incubations, negative selection was performed using anti-rat Dynabeads (Thermo Fisher Scientific). Enriched CD4+ T cells were stained with CellTrace Violet (CTV, Thermo Fisher Scientific) for 10 minutes at 37°C. FACS-purified DC subsets were co-cultured with BALB/c T cells in a 1:5 ratio for 4 days. As a control for homeostatic proliferation, T cells were cultured alone without DCs. Results were expressed as frequency of CTVlow CD4+ T cells.
Anti rat dynabeads
Manufactured by Thermo Fisher Scientific
Sourced in United States
Anti-rat Dynabeads are superparamagnetic beads coated with antibodies specific to rat proteins. They are designed for the rapid and efficient isolation and enrichment of target proteins from rat samples.
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Lab products found in correlation
3 protocols using anti rat dynabeads
1
CD4+ T Cell Proliferation Assay
2
Isolation and Characterization of Primary Mouse Lung Endothelial Cells
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The primary mouse lung endothelial cells (MLEC) were isolated from a 10-week-old, female, Balb/c mice and were cultured as described elsewhere29 (link). Briefly, freshly isolated mouse lung was minced using autoclaved scissors, digested by collagenase I, and filtered through a 70-μm cell strainer. The cell suspension was incubated with anti-rat Dynabeads (Thermo Fisher Scientific, MA, USA, 11035) conjugated with anti-mouse CD31 antibody (BD Biosciences, NJ, USA, 557355). Pooled cells were seeded in a 12-well-plate pre-coated with 0.1% gelatin. Upon reaching confluence, the cells were trypsinized and then incubated with anti-mouse ICAM-2 antibody (BD Biosciences, NJ, USA, 553326) conjugated Dynabeads. Pooled cells were seeded in 12-well-plates pre-coated with 0.1% gelatin. Harvested cells were assessed using tube formation assay, 1,1′-dioctadecyl-3,3,3′,3′-tetramethyl-indocarbocyanine perchlorate Low Density Lipoprotein (DiI-Ac-LDL) uptake, and CD31 gene expression (Fig. S4 ).
3
CD4+ T Cell Proliferation Assay
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Spleens from BALB/c animals were disrupted mechanically. CD4+ T cells were obtained by negative selection using a cocktail of Abs containing the following culture supernatants (produced in house): B220 (RA3-6B2), F4/80 (HB-198), MHC-II (TIB 120), NK1.1 (HB-191) and CD8 (2.43). After supernatant incubations, negative selection was performed using anti-rat Dynabeads (Thermo Fisher Scientific). Enriched CD4+ T cells were stained with CellTrace Violet (CTV, Thermo Fisher Scientific) for 10 minutes at 37°C. FACS-purified DC subsets were co-cultured with BALB/c T cells in a 1:5 ratio for 4 days. As a control for homeostatic proliferation, T cells were cultured alone without DCs. Results were expressed as frequency of CTVlow CD4+ T cells.
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