With a predicted molecular mass of 469 kDa, expression of recombinant DNA-dependent protein kinase catalytic subunit (DNA-PKcs) is challenging. However, DNA-PKcs is relatively abundant in human cells, making it possible to purify the endogenous protein. Here we describe a method to purify DNA-PKcs and its binding partner Ku70/80 from HeLa cells and describe conditions for transfer of DNA-PKcs and other large polypeptides for immunoblotting.
Keywords: DNA-PKcs; HeLa cells; Ku70/80; Protein purification.
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.