Journal: Viruses

Article Title: A Linear Surface Epitope in a Proline-Rich Region of ORF3 Product of Genotype 1 Hepatitis E Virus

doi: 10.3390/v8080227

Figure Lengend Snippet: Genotype 1 VP13 protein reacts with the monoclonal antibody (Mab) in western blotting (WB). ( A ) Detection of purified maltose-binding protein (MBP)-VP13, but not MBP-X or MBP-Helicase. MBP detection was done as a control; ( B ) Detection of only genotype 1 VP13 in lysate of HEK293 cells transiently transfected with YFP-VP13 plasmids or empty vector (EV). Purified MBP-VP13 was included as a positive control. Ab: antibody.

Article Snippet: Antibodies used in the WB were rabbit anti-MBP antibody (Rockland Immunochemicals Inc.), mouse anti-YFP (yellow fluorescent protein) monoclonal antibody (Rockland Immunochemicals Inc.), mouse anti-tubulin monoclonal antibody (Sigma-Aldrich Corp., St. Louis, MO, USA), homemade monoclonal anti-VP13 clone 3C3 (this study) and goat anti-mouse IgG or goat anti-rabbit IgG conjugated with horseradish peroxidase (Rockland Immunochemicals Inc.).

Techniques: Western Blot, Purification, Binding Assay, Control, Transfection, Plasmid Preparation, Positive Control

Journal: Molecular Medicine Reports

Article Title: Comparative proteomic and genomic analyses of Brucella abortus biofilm and planktonic cells

doi: 10.3892/mmr.2019.10888

Figure Lengend Snippet: Differentially expressed proteins in the Brucella abortus A3313 biofilm, identified by MALDI-TOF/TOF-MS analysis.

Article Snippet: Then, the blocked membrane was incubated with sera from primary antibodies [hypothetical protein BAbS19_I16470 (1:1,000; cat. no. orb309412; Biorbyt Ltd.); chaperone protein DnaJ (1:1,000; cat. no. PA3-018; Invitrogen; Thermo Fisher Scientific, Inc.); elongation factor Tu (1:1,000; cat. no. ab210089; Abcam); Chaperonin Cpn60/TCP-1 (1:1,000; cat. no. 3094R-100; BioVision, Inc.); polyprenyl synthetase (1:1,000; cat. no. ab80647; Abcam); periplasmic binding protein (1:1,000; cat. no. M30934-1; Wuhan Boster Biological Technology, Ltd.); enolase (1:1,000; cat. no. sc-271384; Santa Cruz Biotechnology, Inc.); acetyl-CoA carboxylase, a subunit (1:1,000; cat. no. MAB6898; R&D Systems, Inc.); tryptophanyl-tRNA synthetase (1:1,000; cat. no. ab31536; Abcam); aspartate-semialdehyde dehydrogenase (1:1,000; cat. no. EM1708-10a; Jingke Huaxue; exosporium protein B (1:1,000; cat. no. ab92932; Abcam); enoyl-(acyl carrier protein) reductase (1:1,000; cat. no. abx109426; Abbexa Ltd.); Omp16 (1:1,000; cat. no. ab93127; Abcam)] for 2 h at room temperature and then incubated with horseradish peroxidase-labeled secondary antibodies (1:5,000; cat. no. 29139; Invitrogen; Thermo Fisher Scientific, Inc.) in blocking buffer for 1 h at room temperature.

Techniques: Standard Deviation, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Comparative proteomic and genomic analyses of Brucella abortus biofilm and planktonic cells

doi: 10.3892/mmr.2019.10888

Figure Lengend Snippet: Differential expression of mRNAs between Brucella abortus cultured under planktonic or biofilm conditions. The relative expression levels of (A) 18 genes encoding proteins identified by 2-D electrophoresis and (B) 14 genes identified via high-throughput sequencing. 2-D, two-dimensional. *P<0.05 vs. control (planktonic cells). E11-22, hypothetical protein BAbS19_I16470; E15-1, polyprenyl synthetase; E21, ExsB protein; J12, Chaperonin Cpn60TCP-1; E13-1, elongation factor Tu; E20-2, aspartate-semialdehyde dehydrogenase; C24-1, enoyl-(acyl carrier protein) reductase; E12-1, DnaJ, chaperone protein DnaJ; E12-3, isocitrate dehydrogenase; E17, Enolase; E18, Acetyl-CoA carboxylase, alpha subunit; C24-2, putative sulfite oxidase subunit YedY; E11-1, Bacterial protein export chaperone SecB; E13-2, Antifreeze protein, type I; E15-2, Lactatemalate dehydrogenase; E15-5, Phosphoribosylformylglycinamidinecyclo-ligase; E16-3, Periplasmic binding protein; E19-2, tryptophanyl-tRNAsynthetase.

Article Snippet: Then, the blocked membrane was incubated with sera from primary antibodies [hypothetical protein BAbS19_I16470 (1:1,000; cat. no. orb309412; Biorbyt Ltd.); chaperone protein DnaJ (1:1,000; cat. no. PA3-018; Invitrogen; Thermo Fisher Scientific, Inc.); elongation factor Tu (1:1,000; cat. no. ab210089; Abcam); Chaperonin Cpn60/TCP-1 (1:1,000; cat. no. 3094R-100; BioVision, Inc.); polyprenyl synthetase (1:1,000; cat. no. ab80647; Abcam); periplasmic binding protein (1:1,000; cat. no. M30934-1; Wuhan Boster Biological Technology, Ltd.); enolase (1:1,000; cat. no. sc-271384; Santa Cruz Biotechnology, Inc.); acetyl-CoA carboxylase, a subunit (1:1,000; cat. no. MAB6898; R&D Systems, Inc.); tryptophanyl-tRNA synthetase (1:1,000; cat. no. ab31536; Abcam); aspartate-semialdehyde dehydrogenase (1:1,000; cat. no. EM1708-10a; Jingke Huaxue; exosporium protein B (1:1,000; cat. no. ab92932; Abcam); enoyl-(acyl carrier protein) reductase (1:1,000; cat. no. abx109426; Abbexa Ltd.); Omp16 (1:1,000; cat. no. ab93127; Abcam)] for 2 h at room temperature and then incubated with horseradish peroxidase-labeled secondary antibodies (1:5,000; cat. no. 29139; Invitrogen; Thermo Fisher Scientific, Inc.) in blocking buffer for 1 h at room temperature.

Techniques: Quantitative Proteomics, Cell Culture, Expressing, Electrophoresis, Next-Generation Sequencing, Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Comparative proteomic and genomic analyses of Brucella abortus biofilm and planktonic cells

doi: 10.3892/mmr.2019.10888

Figure Lengend Snippet: RT-qPCR identification of the mRNA expression levels of 18 proteins identified in 2-D electrophoresis.

Article Snippet: Then, the blocked membrane was incubated with sera from primary antibodies [hypothetical protein BAbS19_I16470 (1:1,000; cat. no. orb309412; Biorbyt Ltd.); chaperone protein DnaJ (1:1,000; cat. no. PA3-018; Invitrogen; Thermo Fisher Scientific, Inc.); elongation factor Tu (1:1,000; cat. no. ab210089; Abcam); Chaperonin Cpn60/TCP-1 (1:1,000; cat. no. 3094R-100; BioVision, Inc.); polyprenyl synthetase (1:1,000; cat. no. ab80647; Abcam); periplasmic binding protein (1:1,000; cat. no. M30934-1; Wuhan Boster Biological Technology, Ltd.); enolase (1:1,000; cat. no. sc-271384; Santa Cruz Biotechnology, Inc.); acetyl-CoA carboxylase, a subunit (1:1,000; cat. no. MAB6898; R&D Systems, Inc.); tryptophanyl-tRNA synthetase (1:1,000; cat. no. ab31536; Abcam); aspartate-semialdehyde dehydrogenase (1:1,000; cat. no. EM1708-10a; Jingke Huaxue; exosporium protein B (1:1,000; cat. no. ab92932; Abcam); enoyl-(acyl carrier protein) reductase (1:1,000; cat. no. abx109426; Abbexa Ltd.); Omp16 (1:1,000; cat. no. ab93127; Abcam)] for 2 h at room temperature and then incubated with horseradish peroxidase-labeled secondary antibodies (1:5,000; cat. no. 29139; Invitrogen; Thermo Fisher Scientific, Inc.) in blocking buffer for 1 h at room temperature.

Techniques: Expressing, Electrophoresis, Binding Assay