Anti-Human IgG3 Antibody [K16244_1A5]

Various protein samples were run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. K16244_1A5 at 1 µg/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. Human IgG3 band was visualized using ECL Western Blotting Substrate. 

Lane 1: 100 ng of human IgG1 

Lane 2: 100 ng of human IgG2 

Lane 3: 100 ng of human IgG3 

Lane 4: 100 ng of human IgG4 

Lane 5: 100 ng of human IgA 

Lane 6: 100 ng of human IgM 

Lane 7: 100 ng of human IgE 

Lane 8: 100 ng of rat IgG1 

Result: K16244_1A5 can specifically detect IgG3 by Western blotting.

Microtiter wells were coated with K16244_1A5 at 3 µg/mL as the capture antibody. IgG3 was used as the antigen. Peroxidase conjugated mouse anti-IgG3 monoclonal antibody (K06089_8E4) was used as the detection antibody. 

Result: K16244_1A5 and K06089_8E4 can be used as a matched antibody pair to detect and quantify the concentration of IgG3.

IHC-P analysis of human tonsil tissue by anti-IgG3 antibody (K16244_1A5). 

IHC-P was performed using sections of the formalin-fixed paraffin-embedded human tonsil tissue. Antigen was retrieved through addition of boiling Tris/EDTA buffer pH 9 in a pressure cooker for 3 min. Endogenous peroxidase activity was quenched by incubating the sections with 3% H2O2 for 30 min at room temperature. The sections were then incubated with anti-IgG3 primary antibody (K16244_1A5) at 0.8 µg/mL at room temperature for 1 h. Poly-peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. Diaminobenzidine was used as the chromogen. The section was counterstained with hematoxylin. 

Result: Germinal center cells are positively stained at cytoplasm.

Microtiter wells were coated with human immunoglobulins with various isotypes. K16244_1A5 was used as primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. 

Result: K16244_1A5 does not cross-react with human IgG1, IgG2, IgG4, IgA, IgM and IgE.