For activation of polyclonal CD8+ T cells, T cells were isolated from the spleen and inguinal lymph nodes of C57BL/6 mice (Jackson 000664) using the Dynabeads Untouched CD8+ T cell kit. T cells were activated with plate-bound anti-CD3 (2C11, 3 μg/mL, 3:1000 dilution) and anti-CD28 (19.51, 1 μg/mL, 1:500 dilution) for 48 h in RPMI-1640 media containing 10% FBS, 2 mM L-glutamine, 50 μM β-ME, and 10 ng/mL IL-2 (Peprotech). For activation of OT-I transgenic CD8+ T cells, single cell suspensions were generated from spleens and inguinal lymph nodes of OT-I mice (Jackson 003831). Cells were cultured at 10 × 106 cells per mL in the presence of 1 μM SIINFEKL peptide (Invivogen) + for 48 h in RPMI-1640 media containing 10% FBS, 2 mM L-glutamine, 50 μM β-ME, and 10 ng/mL IL-2 (Peprotech).
Dynabeads untouched cd8 t cell kit
Manufactured by Jackson ImmunoResearch
The Dynabeads Untouched CD8+ T cell kit is a magnetic bead-based cell separation system designed to isolate untouched CD8+ T cells from a sample. The kit utilizes a combination of antibodies and magnetic beads to negatively select for CD8+ T cells, leaving them in their native, unmodified state.
Automatically generated - may contain errors
2 protocols using dynabeads untouched cd8 t cell kit
1
Polyclonal and OT-I CD8+ T cell activation
2
Polyclonal and OT-I CD8+ T cell activation
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For activation of polyclonal CD8+ T cells, T cells were isolated from the spleen and inguinal lymph nodes of C57BL/6 mice (Jackson 000664) using the Dynabeads Untouched CD8+ T cell kit. T cells were activated with plate-bound anti-CD3 (2C11, 3 μg/mL, 3:1000 dilution) and anti-CD28 (19.51, 1 μg/mL, 1:500 dilution) for 48 h in RPMI-1640 media containing 10% FBS, 2 mM L-glutamine, 50 μM β-ME, and 10 ng/mL IL-2 (Peprotech). For activation of OT-I transgenic CD8+ T cells, single cell suspensions were generated from spleens and inguinal lymph nodes of OT-I mice (Jackson 003831). Cells were cultured at 10 × 106 cells per mL in the presence of 1 μM SIINFEKL peptide (Invivogen) + for 48 h in RPMI-1640 media containing 10% FBS, 2 mM L-glutamine, 50 μM β-ME, and 10 ng/mL IL-2 (Peprotech).
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