alexa fluor 594 goat anti mouse igg  (Thermo Fisher)

Journal: Nature Communications

Article Title: Prothymosin ? overexpression contributes to the development of pulmonary emphysema

doi: 10.1038/ncomms2906

Figure Lengend Snippet: Overexpression of ProT increases protein acetylation in emphysematous lungs of mice and patients. ( a ) Immunohistochemistry (left) and quantitation (right) of acetylated-lysine (acetyl-K) in lung tissues of 4-day-old ProT HZ, HET and NT mice. ( b ) A positive correlation ( P =0.0012; Pearson’s correlation coefficient) between the levels of ProT immunoreactivity and those of acetylated-lysine immunoreactivity. ( c ) Immunohistochemistry (upper) and quantitation (lower) of acetyl-K in lung tissues from patients with emphysema of varying severity and non-COPD individuals. Values shown are immunoreactive levels in individual specimens; horizontal bars represent the mean. ( d ) Immunofluorescence microscopy. Lung tissues from patients with severe emphysema and from non-COPD individuals were incubated with mouse monoclonal antibody against ProT and rabbit polyclonal antibody against acetyl-K followed by Alexa Fluor 594-conjugated goat anti-mouse IgG and FITC-conjugated goat anti-rabbit IgG. Nuclei were counterstained with DAPI. Fluorescence signals for ProT (red), acetyl-K (green) and the nucleus (blue) were examined by fluorescence microscopy. ( e ) A positive correlation ( P

Article Snippet: For immunofluorescence, the cells were incubated with ProT (clone 2F11) and acetylated-lysine (1:800, Cell Signaling) antibodies and subsequently incubated with Alexa Fluor 594-conjugated goat anti-mouse IgG (Invitrogen) and FITC-conjugated goat anti-rabbit IgG (KPL).

Techniques: Over Expression, Mouse Assay, Immunohistochemistry, Quantitation Assay, Immunofluorescence, Microscopy, Incubation, Fluorescence

Journal: Nature Communications

Article Title: Prothymosin ? overexpression contributes to the development of pulmonary emphysema

doi: 10.1038/ncomms2906

Figure Lengend Snippet: Overexpression of ProT increases NF-κB acetylation in emphysematous lungs of mice and patients. ( a – c ) Immunohistochemical detection of acetyl-NF-κB p65 in lung tissues from 4-day-old ProT HZ, HET and NT mice ( a ), from patients with emphysema of varying severity and non-COPD individuals ( b ) and from FVB mice that had been intratracheally injected with lentiviral vectors expressing ProT or luciferase (Luc) shRNA followed by intraperitoneal injections of CSE or saline twice a week for 6 weeks ( c ). Note that positive nuclear staining for acetyl-NF-κB p65 in the lung epithelium of ProT transgenic mice was observed ( a ) and that higher levels of acetyl-NF-κB p65 accumulation were detected in patients with more severe emphysema ( b ). The boxed areas in upper panels (original magnification × 400; Scale bar=20 μm) are magnified and shown in lower panels. ( d , e ) Immunofluorescence microscopy. Lung tissues from ProT HET and NT mice that had been injected intraperitoneally with CSE or saline twice a week for 6 weeks ( d ) and mouse lung epithelial MLE 12 cells that had been transduced with ProT and GFP by lentiviral vectors Lenti-ProT and Lenti-GFP, respectively, and then treated with CSE or saline ( e ) were incubated with mouse monoclonal antibody against ProT and rabbit polyclonal antibody against NF-κB p65 or acetyl-NF-κB p65 (Lys310) followed by Alexa Fluor 594-conjugated goat anti-mouse IgG and Alexa Fluor 488-conjugated goat anti-rabbit IgG. Nuclei were counterstained with DAPI. Fluorescence signals for ProT (red), NF-κB p65 (green), acetyl-NF-κB p65 (green) and the nucleus (blue) were examined by fluorescence microscopy. Scale bar=10 μm; original magnification × 400. Results are representative of three independent experiments.

Article Snippet: For immunofluorescence, the cells were incubated with ProT (clone 2F11) and acetylated-lysine (1:800, Cell Signaling) antibodies and subsequently incubated with Alexa Fluor 594-conjugated goat anti-mouse IgG (Invitrogen) and FITC-conjugated goat anti-rabbit IgG (KPL).

Techniques: Over Expression, Mouse Assay, Immunohistochemistry, Injection, Expressing, Luciferase, shRNA, Staining, Transgenic Assay, Immunofluorescence, Microscopy, Transduction, Incubation, Fluorescence

Journal: Molecular and Cellular Biology

Article Title: The Lysyl Oxidase Propeptide Interacts with the Receptor-Type Protein Tyrosine Phosphatase Kappa and Inhibits ?-Catenin Transcriptional Activity in Lung Cancer Cells ▿

doi: 10.1128/MCB.01426-10

Figure Lengend Snippet: LOX-PP leads to relocalization to the lysosome and degradation of PIC. (A) H1299 cells were cotransfected with GFP-PIC and a V5-tagged LOX-PP construct or EV DNA, as control. For lysosome staining, a LAMP2 antibody and anti-mouse Alexa Fluor 555-IgG antibody

Article Snippet: The following antibodies were from Invitrogen: secondary anti-mouse IgG–Alexa Fluor 594 (catalog number ), secondary anti-mouse IgG–Alexa Fluor 555 (catalog number ), anti-green fluorescent protein (anti-GFP) (catalog number A-6455), and anti-V5 (catalog number R960-25).

Techniques: Construct, Staining

Journal: Molecular and Cellular Biology

Article Title: The Lysyl Oxidase Propeptide Interacts with the Receptor-Type Protein Tyrosine Phosphatase Kappa and Inhibits ?-Catenin Transcriptional Activity in Lung Cancer Cells ▿

doi: 10.1128/MCB.01426-10

Figure Lengend Snippet: LOX-PP reduces the nuclear levels of the PIC isoform in lung cancer cells. H1299 cells were cotransfected with GFP-PIC and a V5-tagged LOX-PP construct or EV DNA as a control. For LOX-PP staining, a V5 antibody and anti-mouse Alexa Fluor 555-IgG secondary

Article Snippet: The following antibodies were from Invitrogen: secondary anti-mouse IgG–Alexa Fluor 594 (catalog number ), secondary anti-mouse IgG–Alexa Fluor 555 (catalog number ), anti-green fluorescent protein (anti-GFP) (catalog number A-6455), and anti-V5 (catalog number R960-25).

Techniques: Construct, Staining

Journal: Journal of Translational Medicine

Article Title: Intratumoral delivery of recombinant vaccinia virus encoding for ErbB2/Neu inhibits the growth of salivary gland carcinoma cells

doi: 10.1186/1479-5876-12-122

Figure Lengend Snippet: In vitro biological activity of immune sera or purified immunoglobulins from rV- neu T vaccinated mice. Panel A : ADCC elicited by sera from 10 8 pfu rV- neu T vaccinated mice. SALTO tumor cells were exposed for 2 hours to sera pooled from rV- neu T or V-wt vaccinated mice and with mononuclear effector cells derived from normal BALB/c spleens at a ratio of 50:1. Results represent average percent cytotoxicity of two independent experiments. *p ≤ 0.01. Panel B : Effects of anti-Neu Igs on SALTO cell proliferation. SALTO tumor cells following serum depletion were incubated in DMEM medium containing 0.2% BSA with or without rV- neu T or V-wt purified Igs. Relative cell numbers of triplicate experiments were determined after incubation of 72 h at 37˚C using a sulforhodamine B based proliferation assay and expressed as percent increase or decrease in comparison to vehicle control (DMEM 0.2% BSA). *p ≤ 0.01. Panel C : p185 Neu receptor down regulation by anti-Neu Igs. SALTO cells were stained with stained with rV- neu T purified Igs and then with goat anti-mouse IgG Alexa fluor-488-conjugated antibody. Original magnification x 500. Expression and phosphorylation of ERK1 and ERK2 after rV- neu T Igs chronically treatment of SALTO tumor cells. Serum depleted SALTO cells were treated with rV- neu T- and V-wt Igs and the ratio between ERK1 and ERK2 and pERK1/pERK2 expression was analyzed by western blotting. Densitometric ratios between the phosphorylated and total levels of each protein are reported. Panel D : Effects of anti-Neu Igs on SALTO cells apoptosis. SALTO cells were plated at 2.5×10 4 cells/well, incubated in DMEM medium containing 0.2% BSA with or without rV- neu T or V-wt purified Igs (10 μg/ml) and stained with a specific anti-cleaved caspase 3 antibody. Staurosporine (1 μM) treatment was used as positive control. Nuclei were counterstained with Hoechst 33342. Original magnification x400.

Article Snippet: Goat anti-mouse IgG Alexa fluor-488-conjugated antibody and goat anti-rabbit IgG Alexa fluor-594-conjugated antibody were purchased from Life Technologies™ Molecular Probes (Oregon, USA).

Techniques: In Vitro, Activity Assay, Purification, Mouse Assay, Derivative Assay, Serum Depletion, Incubation, Proliferation Assay, Staining, Expressing, Western Blot, Positive Control

Journal: PLoS ONE

Article Title: Recombinant Thrombomodulin Protects Mice against Histone-Induced Lethal Thromboembolism

doi: 10.1371/journal.pone.0075961

Figure Lengend Snippet: Extracellular histones cause fatal thromboembolism in mice. (A) The lethal effect of extracellular histones. Mice were intravenously injected with histones (0-80 µg/g, n = 7-12 per group), and survival was analyzed. (B) Histone-induced thrombocytopenia. Numbers of platelets (PLT), red blood cells (RBC), and white blood cells (WBC) in blood 10 min after infusion with histones (0-95 µg/g, n = 3-7 per group, mean ± S.D.) are shown. Data are presented as percentage of the vehicle group (0 µg/g histones). (C) Distribution of DyLight488-labeled platelets and Alexa-Fluor 594-labeled fibrin(ogen) in lung tissue 10 min after infusion with vehicle or 75 µg/g histones. Nuclei were stained with DAPI. Representative images of n = 4. Scale bar = 100 µm. * P

Article Snippet: Frozen sections (4 µm) were probed with anti-fibrinogen/fibrin IgG (Dako Cytomation, Glostrup, Denmark) followed by Alexa-Fluor 594-conjugated goat anti-rabbit IgG (Invitrogen, Carlsbad, CA).

Techniques: Mouse Assay, Injection, Labeling, Staining