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. Author manuscript; available in PMC: 2008 May 31.
Published in final edited form as: J Immunol Methods. 2007 Apr 5;323(1):11–23. doi: 10.1016/j.jim.2007.03.006

Figure 2.

Figure 2

Serum reactivities in mice one week after the second injection of KRN7000:mCD1d:PPD complexes. (A) FACS-based detection of total antibody reactive to JAWS II dendritic cells loaded with KRN7000, iGb3, or DMSO vehicle. Sera were tested at 1:1000 dilution. Results from responding mice (2 of 4 BALB/c, 2 of 3 BALB/c.CD1d−/−) and a pre-immune mouse are shown. The vertical dotted line indicates the upper limit of fluorescence for negative control staining (i.e., minimum level that exceeded the fluorescence of >99% of cells stained with pre-immune serum). Staining with pre-immune serum was similar to staining with the secondary agent alone (data not shown). (B) ELISA-based detection of IgG reactive to platebound KRN7000- or iGb3-loaded mouse CD1d protein, from two samples as indicated. Sera diluted as indicated; y-axes represent specific absorbance values after subtraction of nonspecific binding of serum antibody to buffer-coated wells. Sera from non-responding mice did not show any detectable mCD1d-specific binding above background.