Journal: Virology Journal

Article Title: Classical swine fever virus triggers RIG-I and MDA5-dependent signaling pathway to IRF-3 and NF-?B activation to promote secretion of interferon and inflammatory cytokines in porcine alveolar macrophages

doi: 10.1186/1743-422X-10-286

Figure Lengend Snippet: Expression and nuclear translocation of IRF-3 after CSFV infection in porcine alveolar macrophages. (A) Expression of IRF-3 was measured by Western Blotting with antibodies specific for IRF-3, and the cells were treated as demonstrated in Figure 1 . (B) Indirect immunofluorescence analysis was used to measure cellular localization of IRF-3 in CSFV-infected PAMs. Cells were mock treated, poly (I:C) stimulated, or infected with Shimen isolates at an MOI of 3. At 24 hpi, cells were fixed and the localization of IRF-3 (red) was observed by fluorescence microscope using immunofluorescence stain with anti-IRF-3 and QDs-SA 605-conjugated and biotinylated secondary antibodies. Nuclei were stained with DAPI. Bar, 10 μm. The experiment was repeated three times and the figure shows a representative experiment. An asterisk indicates a statistically significant difference from uninfected cells, * P

Article Snippet: After washed three times with PBST, cells were further incubated with QDs-SA-conjugated and biotinylated secondary antibodies (1:100, Molecular Probes, Jiayuan Quantum Dots, Wuhai, China) at 37°C for 30 min.

Techniques: Expressing, Translocation Assay, Infection, Western Blot, Immunofluorescence, Fluorescence, Microscopy, Staining

Journal: Virology Journal

Article Title: Classical swine fever virus triggers RIG-I and MDA5-dependent signaling pathway to IRF-3 and NF-?B activation to promote secretion of interferon and inflammatory cytokines in porcine alveolar macrophages

doi: 10.1186/1743-422X-10-286

Figure Lengend Snippet: Protein expression and nuclear translocation of NF- κB after CSFV infection in porcine alveolar macrophages. (A) Expression of NF-κB was measured by Western Blotting with antibodies specific for NF-κB, the cells were treated as demonstrated in Figure 1 . (B) Indirect immunofluorescence analysis was used to measure cellular localization of NF-κB in CSFV-infected PAMs. Cells were treated as indicated in Figure 3 . At 24 hpi, cells were fixed and the localization of NF-κB (red) was observed by fluorescence microscope using immunofluorescence stain with anti-NF-κB/p65 and QDs-SA 605-conjugated biotinylated secondary antibodies. Nuclei were stained with DAPI. Bar, 10 μm. Representative results are shown of one of three separate experiments. An asterisk indicates a statistically significant difference from uninfected cells, * P

Article Snippet: After washed three times with PBST, cells were further incubated with QDs-SA-conjugated and biotinylated secondary antibodies (1:100, Molecular Probes, Jiayuan Quantum Dots, Wuhai, China) at 37°C for 30 min.

Techniques: Expressing, Translocation Assay, Infection, Western Blot, Immunofluorescence, Fluorescence, Microscopy, Staining

Journal: Experimental and Therapeutic Medicine

Article Title: Biotin-avidin mediates the binding of adipose-derived stem cells to a porous β-tricalcium phosphate scaffold: Mandibular regeneration

doi: 10.3892/etm.2015.2961

Figure Lengend Snippet: Comparison of the morphology of Bio-ADSCs on Avi-β-TCP and untreated ADSCs on β-TCP. Scanning electron microscopy micrographs of (A) ADSCs/β-TCP and (B) Bio-ADSCs/Avi-β-TCP constructs following 7 day incubation (magnification, ×1,200). ADSCs, adipose-derived stem cells; β-TCP, β-tricalcium phosphate; Bio-ADSCs, biotinylated ADSCs; Avi-β-TCP, avidin-coated β-TCP.

Article Snippet: After rinsing in PBS three times, the tissue sections were incubated with biotinylated secondary antibody (1:500; Z0420; DAKO, Glostrup, Denmark) at room temperature for 1 h. Peroxidase activity was visualized using 0.05% diaminobenzidine and 0.03% H2O2 in PBS.

Techniques: Electron Microscopy, Construct, Incubation, Derivative Assay, Avidin-Biotin Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Biotin-avidin mediates the binding of adipose-derived stem cells to a porous β-tricalcium phosphate scaffold: Mandibular regeneration

doi: 10.3892/etm.2015.2961

Figure Lengend Snippet: (A and B) Hematoxylin and eosin staining and (C and D) immunohistochemical analysis of osteocalcin protein expression at the mandibular defect area in the (A and C) group 1 and (B and D) group 2 rabbits at 4 weeks post-operation (magnification, ×200). The group 1 and group 2 rabbits received the Bio-ADSCs/Avi-β-TCP and Bio-ADSCs/Avi-β-TCP/PRP constructs, respectively. ADSCs, adipose-derived stem cells; β-TCP, β-tricalcium phosphate; Bio-ADSCs, biotinylated-ADSCs; Avi-β-TCP, avidin-coated β-TCP; PRP, platelet-rich plasma.

Article Snippet: After rinsing in PBS three times, the tissue sections were incubated with biotinylated secondary antibody (1:500; Z0420; DAKO, Glostrup, Denmark) at room temperature for 1 h. Peroxidase activity was visualized using 0.05% diaminobenzidine and 0.03% H2O2 in PBS.

Techniques: Staining, Immunohistochemistry, Expressing, Construct, Derivative Assay, Avidin-Biotin Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Biotin-avidin mediates the binding of adipose-derived stem cells to a porous β-tricalcium phosphate scaffold: Mandibular regeneration

doi: 10.3892/etm.2015.2961

Figure Lengend Snippet: Three-dimensional computed tomography images of mandibular defect reconstruction in the (A) group 1 and (B) group 2 rabbits at 4 weeks post-operation (red arrow). The group 1 and group 2 rabbits received the Bio-ADSCs/Avi-β-TCP and Bio-ADSCs/Avi-β-TCP/PRP constructs, respectively. ADSCs, adipose-derived stem cells; β-TCP, β-tricalcium phosphate; Bio-ADSCs, biotinylated-ADSCs; Avi-β-TCP, avidin-coated β-TCP; PRP, platelet-rich plasma.

Article Snippet: After rinsing in PBS three times, the tissue sections were incubated with biotinylated secondary antibody (1:500; Z0420; DAKO, Glostrup, Denmark) at room temperature for 1 h. Peroxidase activity was visualized using 0.05% diaminobenzidine and 0.03% H2O2 in PBS.

Techniques: Computed Tomography, Construct, Derivative Assay, Avidin-Biotin Assay

Journal: PLoS Genetics

Article Title: The Highly Prolific Phenotype of Lacaune Sheep Is Associated with an Ectopic Expression of the B4GALNT2 Gene within the Ovary

doi: 10.1371/journal.pgen.1003809

Figure Lengend Snippet: B4GALNT2 transferase activity revealed by DBA lectin and KM694 antibody staining in Lacaune sheep ovary. Photomicrographs of ovarian sections from +/+ and L/L ewes and stained either with biotinylated-DBA lectin (500 ng/ml) or KM694 mouse monoclonal antibody (1/1000 dilution). A GalNac treatment (200 µM) was used to compete for DBA staining as specificity control. Sections were counterstained with hematoxylin. A black segment indicates the microscopy magnification scale. GC, granulosa cell layer; TC, theca cell layer; Ant, antral cavity.

Article Snippet: After three 5-min washes in PBS containing 0.1% saponin, sections were incubated for 2 hours at room temperature with the biotinylated secondary antibody (donkey anti-goat, Santa Cruz Biotechnology; donkey anti-mouse, Jackson ImmunoResearch Laboratories) diluted 1∶800 in PBS containing 0.1% saponin and 0.1% BSA, and washed thereafter.

Techniques: Activity Assay, Staining, Microscopy

Journal: PLoS Genetics

Article Title: The Highly Prolific Phenotype of Lacaune Sheep Is Associated with an Ectopic Expression of the B4GALNT2 Gene within the Ovary

doi: 10.1371/journal.pgen.1003809

Figure Lengend Snippet: Western immunoblotting analysis of B4GALNT2 transferase activity in Lacaune sheep granulosa cells and follicular fluids. Granulosa cell protein extracts (50 µg) and follicular fluids (200 µg) from +/+ and L/L large antral follicles were precipitated (P) by agarose-DBA lectin or sepharose-protein A-KM694 monoclonal antibody. The resulting purified glycoproteins were separated on SDS-PAGE, transferred on nitrocellulose membrane and revealed after blotting (B) using biotinylated-DBA lectin or KM694 monoclonal antibody.

Article Snippet: After three 5-min washes in PBS containing 0.1% saponin, sections were incubated for 2 hours at room temperature with the biotinylated secondary antibody (donkey anti-goat, Santa Cruz Biotechnology; donkey anti-mouse, Jackson ImmunoResearch Laboratories) diluted 1∶800 in PBS containing 0.1% saponin and 0.1% BSA, and washed thereafter.

Techniques: Western Blot, Activity Assay, Purification, SDS Page

Journal: PLoS Genetics

Article Title: The Highly Prolific Phenotype of Lacaune Sheep Is Associated with an Ectopic Expression of the B4GALNT2 Gene within the Ovary

doi: 10.1371/journal.pgen.1003809

Figure Lengend Snippet: B4GALNT2 transferase activity revealed by DBA lectin after in vitro overexpression of B4GALNT2 in ovine granulosa cells. Primary ovine granulosa cells from +/+ small antral follicles were transiently transfected with either the pCDNA-hB4GALNT2 expressing the human form of B4GALNT2 or the empty pCDNA3.1 vector. Twenty-four hours after transfection, cells were stained with biotinylated-DBA lectin (500 ng/ml). Arrows indicated DBA positive staining only in B4GALNT2 transfected cells. Cells were counterstained with hematoxylin. A black bar indicates the microscopy magnification scale.

Article Snippet: After three 5-min washes in PBS containing 0.1% saponin, sections were incubated for 2 hours at room temperature with the biotinylated secondary antibody (donkey anti-goat, Santa Cruz Biotechnology; donkey anti-mouse, Jackson ImmunoResearch Laboratories) diluted 1∶800 in PBS containing 0.1% saponin and 0.1% BSA, and washed thereafter.

Techniques: Activity Assay, In Vitro, Over Expression, Transfection, Expressing, Plasmid Preparation, Staining, Microscopy

Journal: Veterinary World

Article Title: Detection of Toxoplasma gondii in cat’s internal organs by immunohistochemistry methods labeled with-[strept] avidin-biotin

doi: 10.14202/vetworld.2017.1035-1039

Figure Lengend Snippet: Toxoplasma positive immunohistochemistry staining of cat with labeled-(Strept) avidin-biotin method: (a) Necrosis is present in kidney and (b) ileum (arrow) (brown color is indicating the positive result of immunohistochemistry) (scale bar=30 µm).

Article Snippet: Slides were incubated with primary polyclonal anti-T. gondii antibody (rabbit) primary antibody (Abcam, USA) diluted 1/100 at 37°C for 32 min and biotinylated secondary antibody (Abcam, USA).

Techniques: Immunohistochemistry, Staining, Labeling, Avidin-Biotin Assay

Journal: Journal of Clinical Investigation

Article Title: Inhibition of endogenous thioredoxin in the heart increases oxidative stress and cardiac hypertrophy

doi: 10.1172/JCI200317700

Figure Lengend Snippet: The activities of the Ras–Raf-1–ERK pathway, p38-MAPK, and p46/p54–JNK1 were examined in Tg-DN-Trx1 mice and NTg littermates. ( a ) Some mice were subjected to aortic banding for 2 weeks. Activities of p42/p44–ERK ( a ), Raf-1 ( b ), p38-MAPK ( e ), and p46/p54–JNK1 ( f ) were determined using anti-phosphospecific Ab’s. The same filter was reprobed with respective non-phosphospecific Ab. ( e ) Cell extracts prepared from cardiac myocytes overexpressing mammalian sterile 20–like kinase 1 (Mst1) were used as positive control (P/C). The gel picture is representative of three to ten experiments in each immunoblot analysis. ( c ) The activity of Ras was determined using the Ras-binding domain in Raf-1 coupled with agarose. ( d ) S-thiolation of Ras was determined as described in Methods. COS-7 cells grown in 60-mm dishes were transfected with the indicated expression plasmids (2 μg). Cells were incubated with biotinylated cysteine (0.5 mM), and S-thiolated proteins were isolated by streptavidin-Sepharose. Samples were subjected to immunoblot analyses with anti-Ras Ab. AS-Trx1, antisense Trx1.

Article Snippet: After washing, biotinylated secondary Ab (anti-mouse IgG; BD Pharmingen) was applied for 1 hour, followed by streptavidin-HRP (BD Pharmingen) for 30 minutes at room temperature.

Techniques: Mouse Assay, Positive Control, Activity Assay, Binding Assay, Transfection, Expressing, Incubation, Isolation

Journal: The Journal of Biological Chemistry

Article Title: WNK2 Kinase Is a Novel Regulator of Essential Neuronal Cation-Chloride Cotransporters *

doi: 10.1074/jbc.M111.222893

Figure Lengend Snippet: Effect of WNK2 and WNK3 on the protein level and surface expression of NKCC1 and KCC4. Western blot analysis of the total and biotinylated fraction of proteins extracted from oocytes injected with NKCC1 ( A ) or KCC4 ( B ) cRNA in the absence or presence of WNK2 or WNK3 cRNA. C and D depict the results of the functional expression performed the same day using oocytes from the same batch for NKCC1 or KCC4, respectively, expressed as bumetanide-sensitive ( C ) or chloride-dependent ( D ) 86 Rb + uptake.

Article Snippet: Following washes with PBS, they were incubated with biotinylated secondary antibodies (Jackson Laboratories, 1:250 dilution) for 2 h at room temperature, washed in PBS, and then incubated with Vectastain ABC Elite solution (Vector Laboratories, Burlingame, CA) for 2 h. Sections were developed using 0.05% DAB (pH 7.4), 0.2% glucose, 0.01% nickel ammonium sulfate, 0.04% ammonium chloride, and 8 mg/ml glucose oxidase, and then rinsed, mounted onto glass slides, allowed to dry, dehydrated, and coverslipped.

Techniques: Expressing, Western Blot, Injection, Functional Assay

Journal: Oncotarget

Article Title: Point-of-care test for cervical cancer in LMICs

doi: 10.18632/oncotarget.7709

Figure Lengend Snippet: Scheme of enzyme-enhanced LFIC for VCP detection Biotinylated gold nanoparticles tethered with streptavidin-bearing HRP at the capture site in the lateral flow strip to generate a signal upon interaction with TMB.

Article Snippet: This was followed by the application of biotinylated goat anti-mouse secondary antibody (GAM IgG) conjugated to HRP (Dakocytomation, Carpinteria, CA).

Techniques: Flow Cytometry, Stripping Membranes