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Purification of DNA-Dependent Protein Kinase Catalytic Subunit (DNA-PKcs) from HeLa Cells.

Authors :
Lee L
Yu Y
Lees-Miller SP
Source :
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2444, pp. 227-241.
Publication Year :
2022

Abstract

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.<br /> (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Details

Language :
English
ISSN :
1940-6029
Volume :
2444
Database :
MEDLINE
Journal :
Methods in molecular biology (Clifton, N.J.)
Publication Type :
Academic Journal
Accession number :
35290641
Full Text :
https://doi.org/10.1007/978-1-0716-2063-2_14