mouse monoclonal anti nectin 1 antibody  (Novus Biologicals)

Journal: Molecular Vision

Article Title: HSV-1 infection of human corneal epithelial cells: Receptor-mediated entry and trends of re-infection

doi:

Figure Lengend Snippet: Immunofluorescence imaging of receptors on HCE cell membrane. Images shown were taken using the FITC filter of confocal microscope (Leica SP20). Cells were blocked for 90 min, washed, and then either mock treated with buffer alone ( B , D , F ) or treated with primary antibodies for Nectin-1 ( A ), HVEM ( C ), and PILR-alpha ( E ). Images were taken after the incubation of HCE cells with FITC-conjugated secondary antibodies. Staining of cells with green demonstrate receptor expression.

Article Snippet: Anti-nectin-1 antibodies used were monoclonal mouse anti-nectin-1 (37–5900; Zymed Laboratories, San Francisco, CA).

Techniques: Immunofluorescence, Imaging, Microscopy, Incubation, Staining, Expressing

Journal: Molecular Vision

Article Title: HSV-1 infection of human corneal epithelial cells: Receptor-mediated entry and trends of re-infection

doi:

Figure Lengend Snippet: Flow cytometry analysis of cell-surface receptor expression. Expression was detected by Fluorescence-activated cell sorter (FACS) analysis. Cells were treated with primary antibodies to Nectin-1 ( A ), HVEM ( B ), or PILR-alpha ( C ). HCE cells stained only with FITC-conjugated secondary antibody were used as background controls and are shown as the dark gray in the figure.

Article Snippet: Anti-nectin-1 antibodies used were monoclonal mouse anti-nectin-1 (37–5900; Zymed Laboratories, San Francisco, CA).

Techniques: Flow Cytometry, Cell Surface Receptor Assay, Expressing, Fluorescence, Staining

Journal: Molecular Vision

Article Title: HSV-1 infection of human corneal epithelial cells: Receptor-mediated entry and trends of re-infection

doi:

Figure Lengend Snippet: Antibodies to the major receptors block HSV-1 entry. Monolayers of cells plated in 96-well culture dishes were incubated with serial dilutions of primary antibodies to Nectin-1, HVEM, and PILR-α for 2 h. Cells were then exposed to HSV-1(KOS)gL86 virus and viral entry was measured 6 h post-infection using a spectrophotometer at an OD of 410 nm. The ratio of entry between cells treated and those untreated are reported as percent blocking. Combinations of antibodies that block these receptors were also studied. Data shown are the means of triplicate determinations (±SD). Anti-nectin-1 ( A ), anti-HVEM ( B ), anti-PILR-α ( C ), a combination of anti-nectin-1 and anti-HVEM antibodies ( D ) and a combination of all three antibodies ( E ).

Article Snippet: Anti-nectin-1 antibodies used were monoclonal mouse anti-nectin-1 (37–5900; Zymed Laboratories, San Francisco, CA).

Techniques: Blocking Assay, Incubation, Infection, Spectrophotometry

Journal: Molecular Vision

Article Title: HSV-1 infection of human corneal epithelial cells: Receptor-mediated entry and trends of re-infection

doi:

Figure Lengend Snippet: Cells demonstrate a decreased level of Nectin-1 receptor expression after primary infection. Monolayers of HCE cells plated on a 6-well tissue culture dish were treated with a viral dose at an MOI of 0.01 and incubated for 48 h at 37 °C. Cells were then treated with MTT to distinguish live from dead cells. FACS was then performed to test levels of Nectin-1 on living infected cells tagged with red fluorescent protein. HCE cells untreated with a viral dose were used as a control and are shown as the green shade.

Article Snippet: Anti-nectin-1 antibodies used were monoclonal mouse anti-nectin-1 (37–5900; Zymed Laboratories, San Francisco, CA).

Techniques: Expressing, Infection, Incubation

Journal: eLife

Article Title: SARS-CoV-2 host-shutoff impacts innate NK cell functions, but antibody-dependent NK activity is strongly activated through non-spike antibodies

doi: 10.7554/eLife.74489

Figure Lengend Snippet:

Article Snippet: Antibody , Mouse monoclonal anti-Nectin-1 , Biolegend , Cat No. 340,402 , Flow cytometry (1:200).

Techniques: Flow Cytometry, Magnetic Beads

Journal: Viruses

Article Title: A Nectin1 Mutant Mouse Model Is Resistant to Pseudorabies Virus Infection

doi: 10.3390/v14050874

Figure Lengend Snippet: Characterization of nectin1 (F129A) mutant mice. ( A ) Gene targeting strategy to introduce F129A mutation in mouse nectin1. Sequences in red letters are gRNA-recognized target harboring F129A mutation (underlined sequences). ( B ) Sequences of nectin1 mutation site in homozygous, heterozygous, and WT mice. The underlined sequences are F129 mutation sites. ( C ) Nectin1 expression status in brain tissues of homozygous, heterozygous, and WT mice. ( D , E ) Growth curves of male ( D ) and female ( E ) F129A mutant mice. Female homozygous mutant mice had significantly reduced body weight compared with WT littermates at five, six, seven, and eight weeks. Data are presented in mean ± standard deviation. p -values represent homozygous versus WT and were analyzed with one-way ANOVA followed by Dunnett’s multiple comparison test. ssODN, single-stranded oligodeoxynucleotides; Homo, homozygous nectin1 (F129A) mutant mice; Hetero, heterozygous nectin1 (F129A) mutant mice; WT, wild-type mice.

Article Snippet: Subsequently, the separated protein in gel was transferred onto the PVDF membrane, which was then blocked in 5% milk and incubated with anti-nectin1 mouse monoclonal antibody (sc-21722, Santa Cruz Biotechnology, Dallas, TX, USA) at 4 °C overnight.

Techniques: Mutagenesis, Introduce, Expressing, Standard Deviation, Comparison

Journal: Viruses

Article Title: A Nectin1 Mutant Mouse Model Is Resistant to Pseudorabies Virus Infection

doi: 10.3390/v14050874

Figure Lengend Snippet: Viral challenge experiment 1 in nectin1 (F129A) mutant mice. ( A ) Experimental groups and virus dose for PRV challenge in mice. ( B ) Typical symptoms present in PRV-infected mice. A WT mouse showed body incoordination and severe wound in the neck because of scratching of the viral injection site at 60 h after challenge (left), whereas no symptoms were observed in nectin1 (F129A) homozygous mutant mouse (right). ( C ) Survival curve for nectin1 (F129A) homozygous mutant and WT mice after PRV challenge within 96 h. Four mice in each group were used for analysis of survival curve. The other three mice in each group were sacrificed to analyze viral infection level in various tissues at 36 h. ( D ) PRV viral loads at 36 h and 96 h in brain and serum by qPCR quantification of PRV gE gene. All data are presented in mean ± standard deviation. Statistically significant differences between mutant and WT groups are indicated by p -values, analyzed with Gehan–Breslow–Wilcoxon test and unpaired t -test for survival curve and viral copies, respectively. Homo, homozygous nectin1 (F129A) mutant mice; WT, wild-type mice.

Article Snippet: Subsequently, the separated protein in gel was transferred onto the PVDF membrane, which was then blocked in 5% milk and incubated with anti-nectin1 mouse monoclonal antibody (sc-21722, Santa Cruz Biotechnology, Dallas, TX, USA) at 4 °C overnight.

Techniques: Mutagenesis, Virus, Infection, Injection, Standard Deviation

Journal: Viruses

Article Title: A Nectin1 Mutant Mouse Model Is Resistant to Pseudorabies Virus Infection

doi: 10.3390/v14050874

Figure Lengend Snippet: Viral challenge experiment 2 in nectin1 (F129A) mutant mice. ( A ) Experimental groups and virus dose for PRV challenge. ( B ) Survival curve for nectin1 (F129A) homozygous mutant, heterozygous mutant, and WT mice after PRV challenge within 96 h. ( C ) PRV viral loads in brain and serum by qPCR quantification of PRV gE gene. All surviving mice were sacrificed to analyze viral load in brains and serum at 96 h after viral challenge. Data are presented in mean ± standard deviation. Statistically significant differences are indicated by p -values, analyzed with Gehan–Breslow–Wilcoxon test and two-way ANOVA for survival curve and viral copies, respectively. Homo, homozygous nectin1 (F129A) mutant mice; Hetero, heterozygous nectin1 (F129A) mutant mice; WT, wild-type mice.

Article Snippet: Subsequently, the separated protein in gel was transferred onto the PVDF membrane, which was then blocked in 5% milk and incubated with anti-nectin1 mouse monoclonal antibody (sc-21722, Santa Cruz Biotechnology, Dallas, TX, USA) at 4 °C overnight.

Techniques: Mutagenesis, Virus, Standard Deviation

Journal: Viruses

Article Title: A Nectin1 Mutant Mouse Model Is Resistant to Pseudorabies Virus Infection

doi: 10.3390/v14050874

Figure Lengend Snippet: Defect in eye development in nectin1 (F129A) mutant mice. ( A ) Nectin1 (F129A) homozygous mutant mice show bilateral microphthalmia. Representative images of the eyes of nectin1 (F129A) homozygous mutant and age-matched WT mice are shown. ( B ) Histological analysis of the eyes of nectin1 (F129A) homozygous mutant and age-matched WT mice. The absent vitreous body and abnormal lenses can be found in homozygous mutant mice. ( C ) Magnified views of the ciliary body in the boxed areas in the panel ( B ). The ciliary body of WT mice displays the double cell layer structure of the ciliary epithelia composed of pigment (black-colored cells) and non-pigment cells (stained only by eosin in cytoplasm), whereas the homozygous mutant mice have deformed ciliary body, in which the ciliary processes with double cell layer structure of the ciliary epithelia are absent. ( D ) Nectin1 expression levels in eyes and brains of homozygous mutant and WT mice. Tissues of two mice from the same litter for each group were used for western blot assay. Homo, homozygous nectin1 (F129A) mutant mice; WT, wild-type mice; L, lens; V, vitreous body; CP, ciliary processes. Scale bars: 500 μm in ( B ), 50 μm in ( C ).

Article Snippet: Subsequently, the separated protein in gel was transferred onto the PVDF membrane, which was then blocked in 5% milk and incubated with anti-nectin1 mouse monoclonal antibody (sc-21722, Santa Cruz Biotechnology, Dallas, TX, USA) at 4 °C overnight.

Techniques: Mutagenesis, Staining, Expressing, Western Blot

Journal: Molecular Therapy Oncolytics

Article Title: Oncolytic Herpes Simplex Virus Prevents Premalignant Lesions from Progressing to Cancer

doi: 10.1016/j.omto.2019.11.003

Figure Lengend Snippet: Changes in the Staining Pattern of the Adherens Junction Protein Nectin-1 Accompany Dysplasia (A–D) IHC for nectin-1 demonstrates the presence of free nectin-1 in dysplastic (A and B), but not in normal (C and D) mucosal cells. (A and C) are high magnifications of (B and D), respectively. Size bars represent 16 μm for all. H&E, hematoxylin and eosin staining; IHC, immunohistochemistry.

Article Snippet: The sections were then stained with rabbit polyclonal HSV-1 antibody or mouse anti-nectin-1 monoclonal antibody, 5 μg/mL (Zymed Laboratories, San Francisco, CA, USA).

Techniques: Staining, Immunohistochemistry