Recombinant anti-Human CHD4 Monoclonal Antibody (Clone K49015_2C12)

Immunoprecipitation: Immunoprecipitation was performed by incubation of 2.5 ug of K49015_2C12 with Ramos lysate containing 200 ug of total protein. After absorption with Protein G beads, the mixture was run on 6-18% SDS-PAGE and blotted onto nitrocellulose membrane. CHD4 (K49015_12F6) at 1 ug/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: Ramos lysate Lane 2: CHD4 immunoprecipitated from Ramos lysate by K49015_2C12 Lane 3: The same as Lane 2 but KT82 was used as IgG isotype control antibody Result: K49015_2C12 can immunoprecipitate CHD4.

Indirect ELISA96-well microtitre plates were coated with 1 ug/mL of CHD4 protein at 4 °C overnight. Plates were blocked with 2 % BSA in PBS-T (0.1 % Tween) at room temperature for 1 hour. Serial diluted of K49015_2C12 was used as the primary antibody and added into the plates for 1 hour incubation. Plates were washed with PBS-T for three times and then incubated with HRP conjugated Human IgG1 for 1 hour. After washed with PBS-T for three times, the plates were incubated with ABTS for 15 minute. The absorbance (optical density) was read with a plate reader.