Protocol detail

Complement-Dependent Cytotoxicity Assay

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CDC assay was performed as previously described method^{21 (link)}, using CLBL-1/luc cells (5 × 10⁵ in 100 µl per well), either rat IgG_2a (Thermo Fisher Scientific Inc.), 4E1-7, or chimeric antibodies, and LOW-TOX®-H rabbit complement (Cedarlane, Burlington, Canada; final concentration 1:40). Live and dead cells were counted using trypan blue dye exclusion assay. For chimeric antibody experiments, CLBL-1/luc cells (1 × 10⁵ in 100 µl per well) were used instead of CLBL-1 cells, and live and dead cells were counted as described above.

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Mizuno T., Kato Y., Kaneko M.K., Sakai Y., Shiga T., Kato M., Tsukui T., Takemoto H., Tokimasa A., Baba K., Nemoto Y., Sakai O, & Igase M. (2020). Generation of a canine anti-canine CD20 antibody for canine lymphoma treatment. Scientific Reports, 10, 11476.

Publication 2020

rat IgG2a LOW-TOX®-H rabbit complement

Antibodies Antibody Assay Cells Chimeric Igg2a Rabbit Trypan blue

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Variable analysis

independent variables

Rat IgG2a (Thermo Fisher Scientific Inc.)
4E1-7
Chimeric antibodies

dependent variables

Live and dead cells counted using trypan blue dye exclusion assay

control variables

LOW-TOX®-H rabbit complement (Cedarlane, Burlington, Canada; final concentration 1:40)
CLBL-1/luc cells (5 × 10^5 in 100 µl per well) for rat IgG2a, 4E1-7, and chimeric antibody experiments
CLBL-1/luc cells (1 × 10^5 in 100 µl per well) for chimeric antibody experiments

controls

Positive control: Not specified
Negative control: rat IgG2a (Thermo Fisher Scientific Inc.)

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