Journal: Infection and Immunity

Article Title: Identification of Acyloxyacyl Hydrolase, a Lipopolysaccharide- Detoxifying Enzyme, in the Murine Urinary Tract

doi: 10.1128/IAI.72.6.3171-3178.2004

Figure Lengend Snippet: Proximal tubule cells secrete pro-AOAH. (A) Diagram of AOAH structure, showing the conversion of the precursor (pro-AOAH) into mature AOAH. (B) Production of 35 S-AOAH by porcine proximal tubule cells in vitro. Labeled AOAH was immunoprecipitated as described in Methods and studied by SDS-PAGE and autoradiography. M, medium; L, lysate. Antibody +, anti-AOAH; antibody −, preimmune IgG. Only pro-AOAH is seen in the media, whereas the lysate contains both pro-AOAH and mature AOAH.

Article Snippet: Wells were blocked for 1 h at 37°C, samples and recombinant human AOAH standard were incubated at 4°C overnight or at 37°C for 2 h, secondary antibody (E553, purified polyclonal rabbit anti-human AOAH, final concentration 10 μg/ml) was added for 1 h at 37°C, biotin-conjugated goat anti-rabbit IgG (0.075 μg/ml) (Zymed, San Francisco, Calif.) was added for 1 h at 37°C, 2 μg of alkaline phosphatase-conjugated streptavidin (Jackson Laboratories)/ml was incubated for 30 min at 37°C, and the plate was developed with alkaline phosphatase substrate (5 mM Sigma 104 and 0.1 M Sigma alkaline buffer 221 in water).

Techniques: In Vitro, Labeling, Immunoprecipitation, SDS Page, Autoradiography

Journal: Nature immunology

Article Title: Autotaxin, a lysophosphatidic acid-producing ectoenzyme, promotes lymphocyte entry into secondary lymphoid organs

doi: 10.1038/ni1573

Figure Lengend Snippet: Secretion of ATX and HECs and transfected MDCK cells. ( a ) Conditioned medium from isolated HECs was subjected to SDS-PAGE and immunoblotted for ATX and GlyCAM-1 The membrane was stripped and reprobed with normal rabbit IgG as a control. The results shown

Article Snippet: 10 µm thick cryostat sections were fixed in acetone, blocked with 3% BSA in PBS containing 10% goat serum and then stained with anti-ATX as a primary antibody followed by biotin conjugated goat anti-rabbit IgG (Jackson ImmuoResearch Laboratories) and Cy2-streptavidin (Jackson ImmuoResearch Laboratories).

Techniques: Transfection, Isolation, SDS Page

Journal:

Article Title: Serum IgG mediates mucosal immunity against rotavirus infection

doi: 10.1073/pnas.0502437102

Figure Lengend Snippet: Fecal rotavirus-specific IgG titers ( a - c ) and IgA titer ( d - f ) of pigtailed macaques passively immunized with high-titer rotavirus-specific IgG ( a and d ), midtiter rotavirus-specific IgG ( b and e ), or nonimmune serum ( c and f ).

Article Snippet: YK-1 infectivity was determined by an immunoassay for rotavirus antigen using a polyclonal rabbit anti-serum for rhesus rotavirus (RRV) as the primary antibody and a biotin-conjugated goat anti-rabbit IgG (Southern Biotechnology Associates) as the secondary antibody.

Techniques:

Journal:

Article Title: Serum IgG mediates mucosal immunity against rotavirus infection

doi: 10.1073/pnas.0502437102

Figure Lengend Snippet: Rotavirus antigen shedding curves after YK-1 challenge of pigtailed macaques passively immunized with high-titer rotavirus-specific IgG ( a ), midtiter rotavirus-specific IgG ( b ), or nonimmune serum ( c ).

Article Snippet: YK-1 infectivity was determined by an immunoassay for rotavirus antigen using a polyclonal rabbit anti-serum for rhesus rotavirus (RRV) as the primary antibody and a biotin-conjugated goat anti-rabbit IgG (Southern Biotechnology Associates) as the secondary antibody.

Techniques:

Journal:

Article Title: Serum IgG mediates mucosal immunity against rotavirus infection

doi: 10.1073/pnas.0502437102

Figure Lengend Snippet: Serum rotavirus-specific IgG titers of pigtailed macaques passively immunized with high-titer rotavirus-specific IgG ( a ), midtiter rotavirus-specific IgG ( b ), or nonimmune serum ( c ).

Article Snippet: YK-1 infectivity was determined by an immunoassay for rotavirus antigen using a polyclonal rabbit anti-serum for rhesus rotavirus (RRV) as the primary antibody and a biotin-conjugated goat anti-rabbit IgG (Southern Biotechnology Associates) as the secondary antibody.

Techniques:

Journal:

Article Title: Serum IgG mediates mucosal immunity against rotavirus infection

doi: 10.1073/pnas.0502437102

Figure Lengend Snippet: Experimental design: passive immunization of pigtailed macaques with pooled serum with rotavirus-specific IgG of high-titer, midtiter, or non-immune control. Macaques were infused with serum and 18 h later challenged with YK-1.

Article Snippet: YK-1 infectivity was determined by an immunoassay for rotavirus antigen using a polyclonal rabbit anti-serum for rhesus rotavirus (RRV) as the primary antibody and a biotin-conjugated goat anti-rabbit IgG (Southern Biotechnology Associates) as the secondary antibody.

Techniques:

Journal: PLoS ONE

Article Title: The Streptococcus pneumoniae Pilus-1 Displays a Biphasic Expression Pattern

doi: 10.1371/journal.pone.0021269

Figure Lengend Snippet: Pilus-1 expression is regulated at the transcriptional level. A) High and low pilus expressing sub-populations of strains TIGR4 (Clade I), 19F Taiwan 14 (Clade I), OREP4 (Clade I), 6B Finland 14 (Clade II) and 35B SME 15 (Clade III) were labeled with clade specific anti-RrgB antibodies (1∶400 dilution) and FITC anti-mouse IgG secondary antibodies (1∶100 dilution). Pilus-1 expression was then analyzed by flow cytometry (FACS-Calibur). B) Schematic representation of PI-1. C) Log 2 ratio values indicating the PI-1 genes differential expression in High vs. Low pilus expressing sub-populations in the five above mentioned strains, as measured by spotted DNA microarray analysis. The data are measures of relative gene expression during in vitro growth in liquid cultures. The values reported for each gene are the mean of all the spots and their replicates within the array and of two independent experiments (bars represent standard deviations). D) Absolute gene expression levels of PI-1 genes measured for TIGR4 high and low pilus expressing sub-populations by microarray hybridization. Absolute expression levels reported for each gene are the mean of all the spots and their replicates within the array and of two independent experiments (bars represent the obtained standard deviations).

Article Snippet: Depletion of RrgB positive bacteria TIGR4 low pilus expressing bacteria were incubated with rabbit anti-RrgB antibodies (1∶400 dilution) and then with goat anti-rabbit IgG biotin conjugated (Abcam, 1∶1000 dilution) antibodies.

Techniques: Expressing, Labeling, Flow Cytometry, Cytometry, FACS, Microarray, In Vitro, Hybridization

Journal: PLoS ONE

Article Title: The Streptococcus pneumoniae Pilus-1 Displays a Biphasic Expression Pattern

doi: 10.1371/journal.pone.0021269

Figure Lengend Snippet: Pilus components display a biphasic expression pattern. A) TIGR4 bacteria were labeled with anti-RrgA, RrgB clade I, RrgC, BgaA (beta-galactosidase), PhtA (pneumococcal histidine triad protein A) or PspC (pneumococcal surface protein C, also known as CbpA,) primary antibodies (1∶400 dilution), and with FITC anti-mouse IgG secondary antibodies (1∶100 dilution). Bacterial staining was analyzed by flow cytometry (FACS-Calibur). Sera of mice immunized with PBS were used as negative control. B,C) TIGR4 bacteria were processed for immunofluorescence, stained with mouse anti-RrgB antibodies (1∶2000 dilution) (red) and with S. pneumoniae anti-capsular antibodies (Omniserum 1∶2000 dilution) (green). Imaging was performed with a confocal microscope. Scale bar is 4 µm in panel B and 1 µm in panel C.

Article Snippet: Depletion of RrgB positive bacteria TIGR4 low pilus expressing bacteria were incubated with rabbit anti-RrgB antibodies (1∶400 dilution) and then with goat anti-rabbit IgG biotin conjugated (Abcam, 1∶1000 dilution) antibodies.

Techniques: Expressing, Labeling, Staining, Flow Cytometry, Cytometry, FACS, Mouse Assay, Negative Control, Immunofluorescence, Imaging, Microscopy

Journal: PLoS ONE

Article Title: The Streptococcus pneumoniae Pilus-1 Displays a Biphasic Expression Pattern

doi: 10.1371/journal.pone.0021269

Figure Lengend Snippet: Pilus expression ratio is not correlated with serotype, genotype or clade type. Bacteria containing either PI-1 clade I (A), clade II (B) or clade III (C), were labeled with clade specific anti-RrgB antibodies (1∶400 dilution) and FITC anti-mouse IgG secondary antibodies (1∶100 dilution). Pilus-1 expression was then analyzed by flow cytometry (FACS-Calibur).

Article Snippet: Depletion of RrgB positive bacteria TIGR4 low pilus expressing bacteria were incubated with rabbit anti-RrgB antibodies (1∶400 dilution) and then with goat anti-rabbit IgG biotin conjugated (Abcam, 1∶1000 dilution) antibodies.

Techniques: Expressing, Labeling, Flow Cytometry, Cytometry, FACS