Anti-Human IgM Antibody [KT194]

Various samples were run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. KT194 at 1 µg/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. Human IgM band was visualized using ECL Western Blotting Substrate.

Lane 1: 100 ng of human IgA

Lane 2: 100 ng of human IgE 

Lane 3: 100 ng of human IgG

Lane 4: 100 ng of human IgM

Lane 5: 100 ng of mouse IgM 

Result: KT194 can specifically detect human IgM by Western blotting.

IHC-P analysis of human tonsil tissue by anti-human IgM antibody

Immunohistochemistry was performed using sections of the 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 IgM primary antibody (KT194) at 2 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. A tissue section incubated with phosphate-buffered saline followed by incubation with the secondary antibody was used as the background control.

Result: A few germinal center cells and squamous epithelial cells are positively stained at the cytoplasm.

Microtiter wells were coated with human immunoglobulins with various isotypes. KT194 was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody.

Result: KT194 does not cross-react with human IgG1, IgG2, IgG3, IgG4, IgA and IgE.