Journal: Microbiology

Article Title: Immunoreactive Coxiella burnetii Nine Mile proteins separated by 2D electrophoresis and identified by tandem mass spectrometry

doi: 10.1099/mic.0.043513-0

Figure Lengend Snippet: Whole-cell extracts of C. burnetii RSA 439 phase II grown in mouse L929 cell culture and separated by 2D gel electrophoresis on pH 3–10 IEF gradients and 10.5–14 % acrylamide SDS-PAGE gels. C. burnetii seroreactive proteins are listed in Table . Molecular mass standards (kDa) are indicated on the left. (a) Sypro Ruby-stained gel showing seroreactive spots. Immune sera-reactive spots are labelled with numbers alone. Sham sera-reactive spots are labelled with numbers with an N prefix. (b) Immunoblot probed with pooled immune guinea pig serum. Spot numbers with an N prefix were also sham sera-reactive. (c) Immunoblot probed with pooled serum from sham-immunized guinea pigs.

Article Snippet: PVDF membranes were dried overnight, stained with Sypro Ruby blot stain (Bio-Rad) according to the manufacturer's instructions and imaged with Quantity One software on a Pharos FX imager (Bio-Rad), and the blot images were printed.

Techniques: Cell Culture, Two-Dimensional Gel Electrophoresis, Electrophoresis, SDS Page, Staining, Western Blot

Journal: Microbiology

Article Title: Immunoreactive Coxiella burnetii Nine Mile proteins separated by 2D electrophoresis and identified by tandem mass spectrometry

doi: 10.1099/mic.0.043513-0

Figure Lengend Snippet: Whole-cell extracts of C. burnetii RSA 439 phase II grown in mouse L929 cell culture and separated by 2D gel electrophoresis on pH 5–8 IEF gradients and 10.5–14 % acrylamide SDS-PAGE gels. C. burnetii seroreactive proteins are listed in Table . Molecular mass standards (kDa) are indicated on the left. (a) Sypro Ruby-stained gel showing seroreactive spots. Immune sera-reactive spots are labelled with numbers alone. Sham sera-reactive spots are labelled with numbers and an N prefix. (b) Immunoblot probed with pooled immune guinea pig serum. (c) Immunoblot probed with pooled serum from sham-immunized guinea pigs.

Article Snippet: PVDF membranes were dried overnight, stained with Sypro Ruby blot stain (Bio-Rad) according to the manufacturer's instructions and imaged with Quantity One software on a Pharos FX imager (Bio-Rad), and the blot images were printed.

Techniques: Cell Culture, Two-Dimensional Gel Electrophoresis, Electrophoresis, SDS Page, Staining, Western Blot

Journal: Microbiology

Article Title: Immunoreactive Coxiella burnetii Nine Mile proteins separated by 2D electrophoresis and identified by tandem mass spectrometry

doi: 10.1099/mic.0.043513-0

Figure Lengend Snippet: Whole-cell extracts of C. burnetii RSA 493 phase I grown in mouse L929 cell culture and separated by 2D gel electrophoresis on pH 5–8 IEF gradients and 10.5–14 % acrylamide SDS-PAGE gels. C. burnetii seroreactive proteins are listed in Table . Molecular mass standards (kDa) are indicated on the left. (a) Sypro Ruby-stained gel showing seroreactive spots. (b) Immunoblot probed with pooled immune guinea pig serum.

Article Snippet: PVDF membranes were dried overnight, stained with Sypro Ruby blot stain (Bio-Rad) according to the manufacturer's instructions and imaged with Quantity One software on a Pharos FX imager (Bio-Rad), and the blot images were printed.

Techniques: Cell Culture, Two-Dimensional Gel Electrophoresis, Electrophoresis, SDS Page, Staining, Western Blot

Journal: Antibiotics

Article Title: Free-Floating Aggregate and Single-Cell-Initiated Biofilms of Staphylococcus aureus

doi: 10.3390/antibiotics10080889

Figure Lengend Snippet: Schematics of experimental setup for growing biofilm in a flow through system.

Article Snippet: For staining the biofilm components, wheat germ agglutinin alexa fluor 647 (W32466, Fisher Scientific, USA; final concentration, 5 μg/mL), DAPI solution (EN62248, Fisher Scientific, USA; final concentration, 1 μg/mL), and SYPRO ruby biofilm matrix stain (F10318, Fisher Scientific, USA) were used to stain the poly-N-acetylglucosamine (PNAG) or SF-derived hyaluronic acid (HA) residues, extracellular DNA (eDNA), and matrix proteins, respectively.

Techniques:

Journal: Antibiotics

Article Title: Free-Floating Aggregate and Single-Cell-Initiated Biofilms of Staphylococcus aureus

doi: 10.3390/antibiotics10080889

Figure Lengend Snippet: Confocal images of aggregated ( A , B ) and single cells ( C , D ) biofilms for 10 8 CFU/mL. Panel B and D are zoomed images. Biofilms stained with DAPI for eDNA, WGA stain for PNAG or SF-derived HA, and SYPRO for staining proteins contained in the biofilm matrix. Merged images denote the combination of GFP, DAPI, WGA, and SYPRO images.

Article Snippet: For staining the biofilm components, wheat germ agglutinin alexa fluor 647 (W32466, Fisher Scientific, USA; final concentration, 5 μg/mL), DAPI solution (EN62248, Fisher Scientific, USA; final concentration, 1 μg/mL), and SYPRO ruby biofilm matrix stain (F10318, Fisher Scientific, USA) were used to stain the poly-N-acetylglucosamine (PNAG) or SF-derived hyaluronic acid (HA) residues, extracellular DNA (eDNA), and matrix proteins, respectively.

Techniques: Staining, Derivative Assay

Journal: Antibiotics

Article Title: Free-Floating Aggregate and Single-Cell-Initiated Biofilms of Staphylococcus aureus

doi: 10.3390/antibiotics10080889

Figure Lengend Snippet: Confocal images of aggregated ( A , B ) and single cells ( C , D ) biofilms for 10 3 CFU/mL. Panel B and D are zoomed images. Biofilms stained with DAPI for eDNA, WGA stain for PNAG or SF-derived HA, and SYPRO for staining proteins contained in the biofilm matrix. Merged images denote the combination of GFP, DAPI, WGA, and SYPRO images.

Article Snippet: For staining the biofilm components, wheat germ agglutinin alexa fluor 647 (W32466, Fisher Scientific, USA; final concentration, 5 μg/mL), DAPI solution (EN62248, Fisher Scientific, USA; final concentration, 1 μg/mL), and SYPRO ruby biofilm matrix stain (F10318, Fisher Scientific, USA) were used to stain the poly-N-acetylglucosamine (PNAG) or SF-derived hyaluronic acid (HA) residues, extracellular DNA (eDNA), and matrix proteins, respectively.

Techniques: Staining, Derivative Assay

Journal: Antibiotics

Article Title: Free-Floating Aggregate and Single-Cell-Initiated Biofilms of Staphylococcus aureus

doi: 10.3390/antibiotics10080889

Figure Lengend Snippet: Confocal images of aggregated ( A , B ) and single cells ( C , D ) biofilms for 10 8 CFU/mL after 3 h antibiotic treatment. Panel B and D are zoomed images. Biofilms stained with DAPI for eDNA, WGA stain for PNAG or SF-derived HA, and PI for staining dead cells. Merged images denote the combination of DAPI, WGA, GFP, and PI images. GFP-PI is combination of GFP and PI images.

Article Snippet: For staining the biofilm components, wheat germ agglutinin alexa fluor 647 (W32466, Fisher Scientific, USA; final concentration, 5 μg/mL), DAPI solution (EN62248, Fisher Scientific, USA; final concentration, 1 μg/mL), and SYPRO ruby biofilm matrix stain (F10318, Fisher Scientific, USA) were used to stain the poly-N-acetylglucosamine (PNAG) or SF-derived hyaluronic acid (HA) residues, extracellular DNA (eDNA), and matrix proteins, respectively.

Techniques: Staining, Derivative Assay

Journal: Antibiotics

Article Title: Free-Floating Aggregate and Single-Cell-Initiated Biofilms of Staphylococcus aureus

doi: 10.3390/antibiotics10080889

Figure Lengend Snippet: Confocal images of aggregated ( A , B ) and single cells ( C , D ) biofilms for 10 8 CFU/mL after 24 h antibiotic treatment. Panel B and D are zoomed images. Biofilms stained with DAPI for eDNA, WGA stain for PNAG or SF-derived HA, and PI for staining dead cells. Merged images denote the combination of DAPI, WGA, GFP, and PI images. GFP-PI is combination of GFP and PI images.

Article Snippet: For staining the biofilm components, wheat germ agglutinin alexa fluor 647 (W32466, Fisher Scientific, USA; final concentration, 5 μg/mL), DAPI solution (EN62248, Fisher Scientific, USA; final concentration, 1 μg/mL), and SYPRO ruby biofilm matrix stain (F10318, Fisher Scientific, USA) were used to stain the poly-N-acetylglucosamine (PNAG) or SF-derived hyaluronic acid (HA) residues, extracellular DNA (eDNA), and matrix proteins, respectively.

Techniques: Staining, Derivative Assay

Journal: Antibiotics

Article Title: Free-Floating Aggregate and Single-Cell-Initiated Biofilms of Staphylococcus aureus

doi: 10.3390/antibiotics10080889

Figure Lengend Snippet: Confocal images of aggregated ( A , B ) and single cells ( C , D ) biofilms for 10 3 CFU/mL after 3 h antibiotic treatment. Panel B and D are zoomed images. Biofilms stained with DAPI for eDNA, WGA stain for PNAG or SF-derived HA, and PI for staining dead cells. Merged images denote the combination of DAPI, WGA, GFP, and PI images. GFP-PI is combination of GFP and PI images.

Article Snippet: For staining the biofilm components, wheat germ agglutinin alexa fluor 647 (W32466, Fisher Scientific, USA; final concentration, 5 μg/mL), DAPI solution (EN62248, Fisher Scientific, USA; final concentration, 1 μg/mL), and SYPRO ruby biofilm matrix stain (F10318, Fisher Scientific, USA) were used to stain the poly-N-acetylglucosamine (PNAG) or SF-derived hyaluronic acid (HA) residues, extracellular DNA (eDNA), and matrix proteins, respectively.

Techniques: Staining, Derivative Assay

Journal: Antibiotics

Article Title: Free-Floating Aggregate and Single-Cell-Initiated Biofilms of Staphylococcus aureus

doi: 10.3390/antibiotics10080889

Figure Lengend Snippet: Confocal images of aggregated ( A , B ) and single cells ( C , D ) biofilms for 10 3 CFU/mL after 24 h antibiotic treatment. Panel B and D are zoomed images. Biofilms stained with DAPI for eDNA, WGA stain for PNAG or SF-derived HA, and PI for staining dead cells. Merged images denote the combination of DAPI, WGA, GFP, and PI images. GFP-PI is combination of GFP and PI images.

Article Snippet: For staining the biofilm components, wheat germ agglutinin alexa fluor 647 (W32466, Fisher Scientific, USA; final concentration, 5 μg/mL), DAPI solution (EN62248, Fisher Scientific, USA; final concentration, 1 μg/mL), and SYPRO ruby biofilm matrix stain (F10318, Fisher Scientific, USA) were used to stain the poly-N-acetylglucosamine (PNAG) or SF-derived hyaluronic acid (HA) residues, extracellular DNA (eDNA), and matrix proteins, respectively.

Techniques: Staining, Derivative Assay

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Critical roles of specimen type and temperature before and during fixation in the detection of phosphoproteins in breast cancer tissues

doi: 10.1038/labinvest.2015.37

Figure Lengend Snippet: Loss of phosphoprotein signal due to delayed preservation (murine samples). Murine breast cancer specimens were either fixed in standard formalin or cold formalin immediately after resection or after different ischemia time points with storage until preservation at room temperature or under vacuum at 4 °C. Protein levels were either determined by reverse phase protein arrays ( a and b ) or by western blot analysis ( c ) with the depicted antibodies. The reverse phase protein arrays data in ( a ) are shown as normalized signal intensities that were calculated by normalization to total protein (SYPRO Ruby staining). Each bar in the reverse phase protein arrays study reflects n =3 data points, besides setting ‘cold formalin, ischemia room temperature', time point 5 h reflects n =4 and time point 24 h reflects n =1. For the setting ‘standard formalin, the ischemia room temperature' time point 24 h is missing because it was not analyzable. In ( b ), two examples (pAKT and pHER2) are highlighted showing the advantage of cold formalin if specimens are kept at 4 °C during the cold ischemia duration. Data are shown as signal intensities in percent relative to the time point zero. Western blot data were normalized to the reference protein beta-Actin before quantification.

Article Snippet: SYPRO Ruby staining was performed according to the manufacturer's instructions and visualized using an E-BOX VX2 UV Transilluminator (Peqlab, Erlangen, Germany).

Techniques: Preserving, Western Blot, Staining

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Critical roles of specimen type and temperature before and during fixation in the detection of phosphoproteins in breast cancer tissues

doi: 10.1038/labinvest.2015.37

Figure Lengend Snippet: Loss of phosphoprotein signal due to delayed preservation (human samples). Human breast cancer biopsies from six patients were immediately fixed in standard formalin, and matched resection specimens were either fixed in standard formalin or cold formalin. Protein expression was determined by reverse phase protein arrays with depicted antibodies. The reverse phase protein arrays data are depicted as normalized signal intensities, which were calculated by normalization to total protein (SYPRO Ruby staining).

Article Snippet: SYPRO Ruby staining was performed according to the manufacturer's instructions and visualized using an E-BOX VX2 UV Transilluminator (Peqlab, Erlangen, Germany).

Techniques: Preserving, Expressing, Staining