Mouse anti-Human ENO1 Monoclonal Antibody (Clone KT162)

ELISA: Microtiter wells were coated with KT162 at 5 ug/mL as the capture antibody. Human ENO1 was used as the antigen. Peroxidase conjugated mouse anti-human ENO1 monoclonal antibody (KT161) was used as the detection antibody. Result: KT162 and KT161 can be used as a matched antibody pair to detect and quantify the concentration of ENO1.

Immunohistochemistry: IHC-P analysis of colon tissue by ENO1 antibody (KT162). IHC-P was performed using sections of the formalin-fixed paraffin-embedded colon tissue.

Immunoprecipitation: Immunoprecipitation was performed by incubation of 2.5 ug KT162 with MCF-7 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. ENO1 (K06117_9F9) at 1 ug/mL was used as the primary antibody and peroxidase conjugated rabbit anti-mouse IgG (Light chain specific) was used as the secondary antibody. The isotype control antibody was KT82. Lane 1: 15 ug of MCF-7 lysate Lane 2: ENO1 immunoprecipitated from MCF-7 lysate by KT162 Lane 3: The same as Lane 2 but KT82 was used as IgG isotype control antibody Result: KT162 can immunoprecipitate ENO1.

Western Blot: Various protein samples were run on 6-18% SDS-PAGE under reducing conditions and blotted onto PVDF membrane. KT162 at 1 ug/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. ENO1 band was visualized using ECL Western Blotting Substrate. Lane 1: 15 ug of SH-SY5Y lysate Lane 2: 15 ug of U-251MG lysate Lane 3: 15 ug of MCF-7 lysate Lane 4: 15 ug of A-431 lysate Lane 5: 15 ug of HeLa lysate Lane 6: 15 ug of THP-1 lysate Lane 7: 15 ug of Jurkat lysate Lane 8: 15 ug of Ramos lysate Lane 9: 15 ug of human plasma Result: KT162 can detect human ENO1 by Western blotting.