Negative controls included samples run without the primary antibody or with nonimmune serum
2] in methanol for 30 minutes, the sections were incubated with normal nonimmune serum
for 1 hour.
Mouse polyclonal antibodies against JEV and WNV and nonimmune serum
samples were included in the assay.
A role for natural antibody in the pathogenesis of leprosy: antibody in nonimmune serum
mediates C3 fixation to the Mycobacterium leprae surface and hence phagocytosis by human mononuclear phagocytes.
In addition, controls for the specificity of double labeling were performed as follows: 1) using nonimmune serum
as the first layer, and 2) using the labeled antisera without the presence of one or both primary antisera.
Equipment manufacturers, to protect their assays from these effects, include nonimmune serum
or immunoglobulin in their reagents, giving a very low incidence of TSH increase.
Sections were pretreated with nonimmune serum
(normal goat serum, 1: 20, for 15 minutes).
Controls consisted of replacement of the primary anti-serum with nonimmune serum
In contrast, injection of nonimmune serum
under the same conditions led to a low and similar reflectivity from all of the spots, which corresponded to nonspecific binding (background value).
Sections were processed using the procedure described herein or in reaction minus the primary antibody and with nonimmune serum
for the negative control.
Formulation of immunoassays requires minimization of interference from heterophilic antibodies and HAAAs; addition of nonimmune serum
from the animal species that was used to raise the antibodies is effective (12,13).
Another study investigated interferences in a two-site CK-MB assay and showed the broad reactivity of interfering globulins to nonimmune serum
from a diverse range of animals (Table 5) (89).