Anti-Human NUCB2 monoclonal antibody

IHC-P analysis of stomach tissue by NUCB2 antibody.
IHC-P was performed using sections of the formalin-fixed paraffin-embedded stomach 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 NUCB2 antibody at 5 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. A tissue section incubated with phosphate-buffered saline followed by incubation with the secondary antibody was used as the background control.
Result: Glandular cells are positively stained at the cytoplasm.

Immunoprecipitation was performed by incubation of 2.5 µg of CABT-L0441Y with HL-60 lysate containing 200 µg of total protein. After absorption with Protein G beads, the mixture was run on 6-18% SDS-PAGE and blotted onto nitrocellulose membrane. CABT-L0441Y at 1 µg/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG (Light chain specific) was used as the secondary antibody. The isotype control antibody was KT82.
Lane 1: HL-60 lysate
Lane 2: NUCB2 immunoprecipitated from HL-60 lysate by CABT-L0441Y
Lane 3: The same as Lane 2 but KT82 was used as IgG isotype control antibody
Result: CABT-L0441Y can immunoprecipitate NUCB2.

15 µg of HL-60 lysate was run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. CABT-L0441Y at 1 µg/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. NUCB2 band was visualized using ECL Western Blotting Substrate.
Result: CABT-L0441Y can detect NUCB2 by Western blotting.