SDS-PAGE analysis of amylase precipitates. Panel A. Precipitates from 18 h culture supernatants, were boiled in sample buffer. The samples containing 3.5 μg of protein were run on 12% SDS-PAGE, followed by staining with SYPRO red in 7.5% acetic acid solution. Lane 1, supernatant of wild type cells (WT); lane 2, supernatant of AbpA-deficient strain (ST); lane 3, supernatant of AbpB-deficient strain (SE); lane 4, molecular mass standard in kDa (M). Panel B. Precipitates from supernatants containing 1.4 μg of protein were used for Gtf-G activity assay. Samples were run on a 12% gel and stained for GTF activity. Lane 1, supernatant from wild type culture (WT); lane 2, supernatant from AbpA-deficient cells (ST); lane 3, supernatant from AbpB-deficient cells (SE). Panel C. Precipitates from supernatants containing 1.4 mg of total protein were used for Western Blot using polyclonal anti-AbpA antibody. Lane1, 2, and 3, supernatants from wild type (WT), AbpA-deficient (ST), and AbpB-deficient cells (SE), respectively.
Chaudhuri et al. BMC Microbiology 2007 7:60 doi:10.1186/1471-2180-7-60