TY  - JOUR
TI  - Purification and Identification of Novel Host-derived Factors for Influenza Virus Replication from Human Nuclear Extracts
AU  - Sugiyama, Kenji 
AU  - Nagata, Kyosuke 
VL  - 6
IS  - 18
PY  - 2016
DA  - 2016/09/20
SP  - e1934
C1  - Bio-protocol 2016;6:e1934
DO  - 10.21769/BioProtoc.1934
UR  - https://doi.org/10.21769/BioProtoc.1934
AB  - Recently, we identified two host cell-derived proteins as novel stimulatory factors of influenza virus RNA replication process, termed “Influenza virus REplication Factor-2 (IREF-2)”, from human nuclear extracts (NEs) by employing biochemical complementation assays (Sugiyama et al., 2015). Herein, we describe detailed methods for successive procedures for identification and purification of IREF-2, including large-scale suspension culture of HeLa S3 cells, preparation of NEs and separation of IREF-2 by sequential column chromatography steps. This protocol can be modified and used for purification and identification of the other unknown nuclear protein(s) of your interest.
KW  - Nuclear Extracts
KW  - Purification and identification
KW  - Influenza
KW  - Host factor
KW  - IREF-2
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False