Rat anti-Mouse Ctcf Monoclonal Antibody (Clone KT70)

Immunoprecipitation: Immunoprecipitation was performed by incubation of 2.5 ug KT70 with MOLT-4 lysate containing 200 ug total protein. After absorption with Protein G beads, the mixture was run on 6-18% SDS-PAGE and blotted onto nitrocellulose membrane. Anti-CTCF (KT71) at 1 ug/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG (which cross reacts with rat IgG) was used as the secondary antibody. The isotype control antibody was KT82. Lane 1: MOLT-4 lysate Lane 2: CTCF immunoprecipitated from MOLT-4 lysate by KT70 Lane 3: The same as Lane 2 but KT82 was used as IgG isotype control antibody Result: KT70 can immunoprecipitate CTCF.

Western Blot: 15 ug of NIH/3T3 lysate was run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. KT70 at 1 ug/mL was used as the primary antibody and peroxidase conjugated goat anti-rat IgG was used as the secondary antibody. CTCF band was visualized using ECL Western Blotting Substrate. Result: KT70 can detect CTCF by Western blotting.

Western Blot: Various protein samples were run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. KT70 at 1 ug/mL was used as the primary antibody and peroxidase conjugated goat anti-rat IgG was used as the secondary antibody. CTCF band was visualized using ECL Western Blotting Substrate. Lane 1: 15 ug of HeLa lysate Lane 2: 15 ug of Jurkat lysate Lane 3: 15 ug of MOLT-4 lysate Result: KT70 can detect CTCF by Western blotting.