Detection of human IgG4 in monkey serum, using Anti-Human IgG4 Antibody [RM120] (capture antibody) and Anti-Human Ig Light Chain Antibody [RM129] (Biotin) (detection antibody) as a Sandwich ELISA pair. HRP conjugated streptavidin and TMB were used to yield the colorimetric reaction.
Sandwich ELISA using Anti-Human IgG4 Antibody [RM120] as the capture antibody, and Anti-Human Ig Light Chain Antibody [RM129] (Biotin) as the detection antibody, followed by an AP conjugated streptavidin.
Figure 3: Western Blot - Anti-Human IgG4 Antibody [RM120] (A121351)
Western blot of human, mouse, rat, and goat IgG shows Anti-Human IgG4 Antibody [RM120] reacts to human IgG4, in both whole molecule (~150kDa, non-reduced) and heavy chain (~50kDa, reduced) forms. There is no cross reactivity with other isotypes of human IgG, or mouse, rat, or goat IgG.
A titer ELISA using Anti-Human IgG4 Antibody [RM120]. The plate was coated with different amounts of human IgG4. A serial dilution of Anti-Human IgG4 Antibody [RM120] was used as the primary antibody, followed by an alkaline phosphatase conjugated anti-rabbit IgGas the secondary antibody.
ELISA showing Anti-Human IgG4 Antibody [RM120] does not react to monkey IgG. The plate was coated with Rhesus monkey IgG. A serial dilution of Anti-Human IgG4 Antibody [RM120] and a monkey IgG binding antibody (positive control) was used as the detection antibody.