Anti-Human IgG4 Antibody [K06216_14H8]

Various samples were run on 6-18% SDS-PAGE under reducing conditions and blottedonto nitrocellulose membrane. K06216_14H8 at 1 µg/mL was used as the primary antibodyand peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody.

Human IgG4 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: K06216_14H8 can specifically detect human IgG4 by Western blotting.

IHC-P analysis of human tonsil tissue by anti-human IgG4 antibody.

Immunohistochemistry was performed using sections of formalin-fixed paraffin-embedded human tonsil. Antigen was retrieved through addition of boiling Tris/EDTA buffer pH 9 in a pressure cooker for 3 min. Removal of endogenous peroxidase activity was done by incubating the sections with 3% H2O2 for 30 min at room temperature. The sections were then incubated with anti-human IgG4 primary antibody (K06216_14H8) at 1 µg/mL at room temperature for 1 h and 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: A few germinal center cells and non-germinal center cells are positively stained at the cytoplasm.

Microtiter wells were coated with human immunoglobulins with various isotypes. K06216_14H8 was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody.

Result: K06216_14H8 does not cross-react with human IgG1, IgG2, IgG3, IgA, IgM and IgE.