NFκB (p65) DNA-binding activity

Following the applied treatment, nuclear proteins were extracted. By using a TransAM™ ELISA kit (Active Motif, Carlsbad, CA, 43296) the NFκB (p65) DNA-binding activity was tested. For each treatment, 1.0 μg of nuclear extracts were subjected to the binding of p65 to an immobilized consensus sequence in a 96-well plate (1 ×10⁴ cells/well). After the colorimetric reaction, OD value was measured in an ELISA reader at the test wavelength of 450 nm.