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. 2015 Oct 22;8(1):129–140. doi: 10.1080/19420862.2015.1109757

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© 2016 The Author(s). Published with license by Taylor & Francis Group, LLC

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 4. — Optimization of single memory B ELISPOT assays (A) Single B cells were sorted using a number of different flow cytometric staining panels and capture media with either 20% or 40% addition of fetal bovine serum. Cells were cultured for 14 d in the presence of IL-21 and feeder cells expressing huCD40L. Bars (mean with SD) indicate the number of wells out of a 96-well plate that resulted in expression of IgG as measured by a total IgG ELISA. Results are grouped into a high (>1 .000), medium (0.5–0.999) and low (0.100–0.499) optical density readouts. (B) Supernatants from 2 96-well plates for each condition were pooled and assayed for IgG concentration in an ELISA assay.