Anti-HSP90AA1 monoclonal antibody

IHC-P analysis of testis tissue by HSP90AA1 antibody.
IHC-P was performed using sections of the formalin-fixed paraffin-embedded testis. Antigen retrieval was done using 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 HSP90AA1 antibody at 2 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.
Results: Cells in seminiferous ducts are positively stained at cytoplasm and membrane

Immunoprecipitation was performed by incubation of 2.5 µg DCABH-9258 with MCF-7 cell lysate containing 200 µg total protein. After absorption with Protein G beads, the mixture was run on 6-18% SDS-PAGE and blotted onto nitrocellulose membrane. DCABH-9258 at 1 µg/mL was used as the primary antibody and peroxidase- conjugated goat anti-mouse IgG was used as the secondary antibody. The isotype control antibody was KT82.
Lane 1: 7.5 µg of MCF-7 lysate
Lane 2: HSP90AA1 immunoprecipitated from MCF-7 lysate by DCABH-9258
Lane 3: The same as Lane 2 but KT82 was used as IgG isotype control antibody.
Result: DCABH-9258 can immunoprecipitate HSP90AA1.

Various protein samples were run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. DCABH-9258 at 1 µg/mL was used as the primary antibody and peroxidase-conjugated goat anti-mouse IgG was used as the secondary antibody. HSP90AA1 band was visualized using ECL Western Blotting Substrate.
Lane 1: 15 µg of HeLa lysate
Lane 2: 15 µg of MCF-7 lysate
Lane 3: 15 µg of HT-29 lysate
Lane 4: 15 µg of HEK293 lysate
Lane 5: 15 µg of Jurkat lysate
Lane 6: 15 µg of K562 lysate
Result: DCABH-9258 can detect HSP90AA1 by Western blotting.