Cell Isolation Naive Pan T cell

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Protocol tips
T cells were purified by magnetic bead selection using the Miltenyi pan-T cell isolation kit (Miltenyi Biotec, Auburn, CA). CLA− and CLA+ subsets were isolated by subsequent incubation of T cells in biotin anti-human CLA (BD Pharmingen) followed by anti-biotin microbeads and magnetic separation (Miltenyi).
Downstream tips
Positively selected cells were collected as the CLA+-enriched fraction and negatively selected cells were collected as the CLA− subset.
Protocol tips
T cells were isolated (at >95% purity as determined by CD3
expression) via magnetic-activated cell sorting (MACS) using the Naïve Pan T Cell Isolation Kit
per the manufacturer’s instructions (Stemcell Technologies) and activated with a non-specific activation stimuli method
Protocol tips
Pan-T cells were isolated from 30 mL whole blood using human MACSxpress Whole Blood Pan T Cell Isolation Kit (Miltenyi Biotec).
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