CD4+ T cells were isolated from splenocytes of BALB/c mice with anti-CD4 magnetic particles (BD Biosciences). On passing through columns (BD Biosciences), the labeled CD4+ T cells were purified to ~99%. To determine T cell priming capacities, DCs were pretreated with mitomycin C and added at different densities (10:1, 20:1, 50:1 and 100:1) to T cells for 3 days. BrdU ELISA was used to determine T cell proliferation according to the manufacturer’s instructions (Merck Millipore, Billerica, MA, USA). Tregs concentration in the coculture system was quantified by flow cytometry as described above.
Anti cd4 magnetic particles
Manufactured by BD
Anti-CD4 magnetic particles are a lab equipment product used for the isolation and separation of CD4+ T cells from biological samples. The particles are coated with antibodies that specifically bind to the CD4 receptor on the surface of these cells, allowing them to be isolated using a magnetic field.
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4 protocols using anti cd4 magnetic particles
1
CD4+ T Cell Proliferation Assay
2
CD4+ T Cell Isolation and Activation
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Peripheral blood samples were collected from patients and treated with Ficoll-Hypaque density gradient centrifugation. CD4 + T cells were isolated using anti-CD4 magnetic particles (BD Biosciences). For stimulation, CD4 + T cells were subjected to anti-CD3 and anti-CD28 antibodies (BD Biosciences Pharmingen) as described before [8 (link)].
3
Isolation and Activation of CD4+ T Cells
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Peripheral blood samples collected from patients were subjected to Ficoll‐Hypaque density gradient centrifugation. Anti‐CD4 magnetic particles from BD Biosciences were used to isolate CD4+ T cells. Anti‐CD3 and anti‐CD28 antibodies (BD Biosciences) were applied to activate CD4+ T cells as described before.24
4
CD4 T cell Activation Protocol
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CD4 T cells were isolated by magnetic separation using anti-CD4 magnetic particles (BD Pharmingen). Cells were activated after isolation with soluble anti-CD3ε (145-2C11) and anti-CD28 (clone 37.51) (BD Pharmingen) 1 μg/mL each, crosslinked with anti-hamster IgG (Sigma) 4.5 μL/mL. Cells were activated at 1.5 × 106 cells/mL. Cells were activated in a 1:1 mixture of RPMI and DMEM (RDG) supplemented with 10% Fetal Bovine Serum (PEAK), L-Glutamine, Na-Pyruvate, Penicillin/Streptomycin, and 2-mercaptoethanol.
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