Sandwich ELISA using RM122 as the capture antibody (25ng/well), and Biotinylated anti-human light chains ( κ+ λ) antibody RM129 as the detection antibody, followed by an alkaline phosphatase conjugated streptavidin.
ELISA of human immunoglobulins shows RM122 reacts only to human IgEλ from human myeloma plasma and the IgE from human plasma. No cross reactivity with Human IgG, IgM, IgD, or IgA. The plate was coated with 50 ng/well of different immunoglobulins. 200 ng/mL, 50 ng/mL or 10 ng/mL of RM122 was used as the primary antibody. An alkaline phosphatase conjugated anti-rabbit IgG as the secondary antibody.
A titer ELISA using RM122. The plate was coated with different amounts of human IgE. A serial dilution of RM122 was used as the primary antibody. An alkaline phosphatase conjugated anti-rabbit IgG as the secondary antibody.