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. 2020 Sep 4;9(3):45. doi: 10.3390/antib9030045

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Two-step CDC. Reaction of OFA-opsonized Daudi cells in C5-depleted (C5-dpl) NHS in step 1 promotes C3b/iC3b deposition and these cells are killed when they are incubated in either NHS or NHS + EDTA in step 2. (A) C3b/iC3b deposition was assayed with PE mAb 7C12 and is expressed as molecules of equivalent soluble fluorochrome (MESF). In a control to block C3b deposition, cells were opsonized in step 1 with C5-depleted NHS + EDTA; the resulting signal was only 1200 MESF. (B) The percentage of dead cells was determined based on uptake of TO-PRO-3. (A,B) Representative of five similar experiments. (C) Killing of cells under various step 2 conditions. Means and SD are displayed. Representative of three similar experiments [22].