EasySep™ HLA T Cell Enrichment Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Donor lymphocytes isolated as T and B cells (EasySep HLA T Cell Enrichment Kit and EasySep HLA B Cell Enrichment Kit, StemCell Technologies, Vancouver, Canada) were added separately to the wells of a Terasaki plate (1 × 105 1 μL buffer) together with 1 μL recipient sera.	The plate was incubated for 40 minutes at 20°C to 25°C followed by a further 70 minutes after adding 5 μL rabbit complement (Bio-Rad).	Then the samples were visualized with 3 μL of eosin and fixed with 5 μL of formalin. The result was acquired by visual inspection under a phase-contrast microscope.

Upstream tips
Donor lymphocytes isolated as T and B cells (EasySep HLA T Cell Enrichment Kit and EasySep HLA B Cell Enrichment Kit, StemCell Technologies, Vancouver, Canada) were added separately to the wells of a Terasaki plate (1 × 105 1 μL buffer) together with 1 μL recipient sera.
Protocol tips
The plate was incubated for 40 minutes at 20°C to 25°C followed by a further 70 minutes after adding 5 μL rabbit complement (Bio-Rad).
Downstream tips
Then the samples were visualized with 3 μL of eosin and fixed with 5 μL of formalin. The result was acquired by visual inspection under a phase-contrast microscope.

Publication protocol

"CDC-XM
Donor lymphocytes isolated as T and B cells (EasySep HLA T Cell Enrichment Kit and EasySep HLA B Cell Enrichment Kit, StemCell Technologies, Vancouver, Canada) were added separately to the wells of a Terasaki plate (1 × 105 1 μL buffer) together with 1 μL recipient sera. The plate was incubated for 40 minutes at 20°C to 25°C followed by a further 70 minutes after adding 5 μL rabbit complement (Bio-Rad). Then the samples were visualized with 3 μL of eosin and fixed with 5 μL of formalin. The result was acquired by visual inspection under a phase-contrast microscope. The negative control serum was an antibody-negative sample pooled from 15 male blood group AB donors."

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