Affinity purification and Western analysis of Rfa3 and Rfa8 in
Halobacterium sp. NRC-1 and derivative strains. Total
protein samples from cell lysates and fractions binding to a DNA-cellulose
column were electrophoresed on a 12% polyacrylamide-SDS gel, and stained with
Coomassie blue, or transferred to PVDF membrane, and probed with rabbit Rfa3
antibody followed by a secondary goat anti-rabbit antibody-alkaline phosphatase
conjugate and substrate. Panel A contains Halobacterium sp.
NRC-1, panel B contains Halobacterium sp. LH5, panel C contains
Halobacterium sp. LH7a, and panel D contains
Halobacterium sp. NRC-1 (pDRK1). Lanes labeled 1 contain
total cell lysate and lanes labeled 2 contain peak bound fraction from a
DNA-cellulose column, Coomassie stained, and lanes labeled 3 display results of
Western blots.