Journal: PLoS ONE

Article Title: Interleukin-19 Acts as a Negative Autocrine Regulator of Activated Microglia

doi: 10.1371/journal.pone.0118640

Figure Lengend Snippet: (A) Western blotting data for receptors of IL-19. Only microglia express both the IL-20Rα and IL-20Rβ subunits. Neu, neurons; Ast, astrocytes; Mi, microglia. (B) Western blotting data for receptors of IL-19. LPS stimulation did not change the expression levels of IL-19 receptor subunits. Skin was used as the positive control. NT, untreated; LPS, LPS-treated. (C) Western-blotting analysis of STAT3 phosphorylation in microglia treated with 100 ng/ml IL-19. IL-19 induced STAT3 phosphorylation, the main downstream signal of the IL-19 receptor. All quantitative data are expressed as means ± SD (n = 3). *, p < 0.01 vs . 0 min. **, p < 0.001 vs . 0 min.

Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.05% Tween-20 for 1 h at room temperature, and then incubated overnight at 4°C with rabbit anti-IL-20Rα polyclonal antibodies (Merck Millipore, Billerica, MA, USA), rat anti-IL-20Rβ monoclonal antibody (eBioscience, San Diego, CA, USA), mouse anti-STAT3 monoclonal antibody (BD Pharmingen, Franklin Lakes, NJ, USA), rabbit anti-pSTAT3 polyclonal antibodies (Cell Signaling Technology, Danvers, MA, USA), or mouse anti-β-actin monoclonal antibody (Sigma-Aldrich), followed by incubation with horseradish peroxidase—conjugated secondary antibodies (GE Healthcare, Buckingham, UK) for 1 h at room temperature.

Techniques: Western Blot, Expressing, Positive Control, Phospho-proteomics

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Development of a Highly Sensitive Bioluminescent Enzyme Immunoassay for Hepatitis B Virus Surface Antigen Capable of Detecting Divergent Mutants

doi: 10.1128/CVI.00186-13

Figure Lengend Snippet: Characterization of the anti-HBs MAbs and the rabbit polyclonal antibody by the inhibition test. Captured HBsAg on rabbit anti-HBs polyclonal antibody-coated magnetic particles was reacted with each unlabeled antibody. Enzyme-labeled anti-HBs antibody was then added to compare the inhibition by each unlabeled antibody. The percent inhibition is shown as the B/B0 ratio on the y axis, where B is bioluminescent intensity when unlabeled antibody is added, and B0 is the bioluminescent intensity when unlabeled antibody is not added.

Article Snippet: The magnetic particles used in the inhibition test were coated with 0.15 mg/ml of anti-HBs rabbit polyclonal antibodies (Biokit, Barcelona, Spain) and prepared in the same way as the MAbs described above.

Techniques: Inhibition, Labeling

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Development of a Highly Sensitive Bioluminescent Enzyme Immunoassay for Hepatitis B Virus Surface Antigen Capable of Detecting Divergent Mutants

doi: 10.1128/CVI.00186-13

Figure Lengend Snippet: Comparison of the detectability during the late phase of infection (days since first bleed) between the BLEIA and PCR. The seroconversion panel PHM935(B) was used to detect HBsAg by the BLEIA. The measurement results of the HBsAg BLEIA are shown in mIU/ml and were compared against various HBV markers, including HBsAg by CLIA2, anti-HBc, anti-HBs, and HBV DNA by PCR, whose data were obtained from the data sheet included with the seroconversion panel. The horizontal dotted line indicates the cutoff value for both the BLEIA (5 mIU/ml) and PCR (400 copies/ml). ▲, HBV PCR-positive data; △, HBV PCR-negative data; ● HBsAg BLEIA-positive data. Top, HBsAg results by CLIA2, anti-HBc, and anti-HBs are shown as + and −.

Article Snippet: The magnetic particles used in the inhibition test were coated with 0.15 mg/ml of anti-HBs rabbit polyclonal antibodies (Biokit, Barcelona, Spain) and prepared in the same way as the MAbs described above.

Techniques: Infection

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Development of a Highly Sensitive Bioluminescent Enzyme Immunoassay for Hepatitis B Virus Surface Antigen Capable of Detecting Divergent Mutants

doi: 10.1128/CVI.00186-13

Figure Lengend Snippet: Results of the analysis of discrepant and nondiscrepant specimens by BLEIA

Article Snippet: The magnetic particles used in the inhibition test were coated with 0.15 mg/ml of anti-HBs rabbit polyclonal antibodies (Biokit, Barcelona, Spain) and prepared in the same way as the MAbs described above.

Techniques:

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Development of a Highly Sensitive Bioluminescent Enzyme Immunoassay for Hepatitis B Virus Surface Antigen Capable of Detecting Divergent Mutants

doi: 10.1128/CVI.00186-13

Figure Lengend Snippet: Detection of preS1/preS2/S proteins by Western blotting. Electrophoresis was conducted in 10 to 20% polyacrylamide gels. The nondiscrepant specimen (P35; see Table 4) was serially diluted 1:2 with 1× sample buffer. The concentration decreases by half going from left to right. The discrepant specimen (P38; see Table 4) was prepared in the same way. (A) A 10-μg/ml preparation of HRP-labeled anti-HBs rabbit polyclonal antibody was used for detection. (B) A 10-μg/ml preparation of anti-preS1 MAb was used as a primary antibody, and 500-fold-diluted HRP-labeled anti-mouse immunoglobulin (IgG) was used as a secondary antibody. (C) A 10-μg/ml preparation of anti-preS2 MAb was used as a primary antibody, and 500-fold-diluted HRP-labeled anti-mouse IgG was used as a secondary antibody. The large HBs proteins (p39 and gp42), middle HBs proteins (gp33 and gp36), and small HBs (S) proteins (p24 and gp27) are marked with arrows.

Article Snippet: The magnetic particles used in the inhibition test were coated with 0.15 mg/ml of anti-HBs rabbit polyclonal antibodies (Biokit, Barcelona, Spain) and prepared in the same way as the MAbs described above.

Techniques: Western Blot, Electrophoresis, Concentration Assay, Labeling

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Development of a Highly Sensitive Bioluminescent Enzyme Immunoassay for Hepatitis B Virus Surface Antigen Capable of Detecting Divergent Mutants

doi: 10.1128/CVI.00186-13

Figure Lengend Snippet: Results of the analysis of nondiscrepant specimens following protease treatment

Article Snippet: The magnetic particles used in the inhibition test were coated with 0.15 mg/ml of anti-HBs rabbit polyclonal antibodies (Biokit, Barcelona, Spain) and prepared in the same way as the MAbs described above.

Techniques: