Journal: Nucleic Acids Research

Article Title: A WYL transcriptional regulator activates the DNA damage response pathway in Acinetobacter species

doi: 10.1093/nar/gkag250

Figure Lengend Snippet: A C-terminal 3xFLAG tag constitutively activates AciT. ( A ) Representative microscopy images of indicated strains. Scale bars, 10 µm. ( B ) Representative western blot on indicated strains grown with or without 2 µg/ml MMC. Blot was probed with ⍺-RpoA antibody as a loading control and ⍺-FLAG antibody to probe for AciT expression. ( C ) Growth curves of indicated strains grown with indicated concentrations of vancomycin in 96-well plates over 12 h. Arrows indicate concentration used for selection experiments. Each dot shows the mean of three independent, biological replicates, and error bars show SEM.

Article Snippet: Then, 1 μl of overnight cultures were subcultured into 200 μl fresh LB medium with vancomycin (MP Biomedicals) (0 μg/ml, 25 μg/ml, 50 μg/ml, 100 μg/ml, 250 μg/ml) grown in a 96-well plate in a Synergy H1 multimode plate reader (BioTek) utilizing Gen 6 software at 30°C shaking for 16 h. OD 600 was measured every 10 min.

Techniques: Microscopy, Western Blot, Control, Expressing, Concentration Assay, Selection

Journal: Chembiochem : a European journal of chemical biology

Article Title: HER2-and EGFR-specific Affiprobes--Novel Recombinant Optical Probes for Cell Imaging

doi: 10.1002/cbic.200900532

Figure Lengend Snippet: Structure of the Affiprobes. Affiprobe molecules contain a hexahistidine tag at the N-terminus, followed by an Affibody molecule targeting moiety Z(HER2), Z(EGFR), and Z(Taq4:5), flexible 6 amino-acid G3SG2 linker, and a fluorescent protein mCherry or EGFP at the C-terminus.

Article Snippet: Affiprobe overexpression and purification BL21 Star®(DE3) E. coli cells (Invitrogen, Carlsbad, CA), transformed with the Affiprobe plasmids, were grown in 1 L Super Broth media (MP Biomedicals, LLC, OH), supplemented with 100 μg/mL carbenicillin (Sigma-Aldrich, MO) until A 600 reached value 1.0.

Techniques:

Journal: Chembiochem : a European journal of chemical biology

Article Title: HER2-and EGFR-specific Affiprobes--Novel Recombinant Optical Probes for Cell Imaging

doi: 10.1002/cbic.200900532

Figure Lengend Snippet: Affiprobe staining of cells with different HER2 and EGFR phenotype. Cells were incubated with Affiprobes (100 μg/mL) for 1h at 37°C in the presence of Hoechst 33342 (1 μg/mL) Representative confocal images through middle section of the cells were acquired with a Zeiss LSM 510 confocal microscope as described in Experimental section.

Article Snippet: Affiprobe overexpression and purification BL21 Star®(DE3) E. coli cells (Invitrogen, Carlsbad, CA), transformed with the Affiprobe plasmids, were grown in 1 L Super Broth media (MP Biomedicals, LLC, OH), supplemented with 100 μg/mL carbenicillin (Sigma-Aldrich, MO) until A 600 reached value 1.0.

Techniques: Staining, Incubation, Microscopy