muc5ac concentrations  (BlueGene Biotech)

Journal: Frontiers in Immunology

Article Title: Plexin B1 controls Treg numbers, limits allergic airway inflammation, and regulates mucins

doi: 10.3389/fimmu.2023.1297354

Figure Lengend Snippet: Plexin B1 regulates mucin expression in lungs. BALF was evaluated for MUC5AC by ELISA, n=7 mice/group (A) . (B) Lung RNA was isolated from the indicated mice and evaluated for mucin gene expression by qRT-PCR as described in Materials and Methods (n=4-8/group). Data shown as mean ± SEM (A, B) with data statistics calculated using two-way ANOVA with Tukey’s multiple comparison test.

Article Snippet: The Muc5AC concentrations in BALF were determined with the use of ELISA kit NBP2-76704 (Novus Biologicals) according to the manufacturer’s instruction.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Isolation, Gene Expression, Quantitative RT-PCR, Comparison

Journal: Frontiers in Pharmacology

Article Title: Airway Mucins Inhibit Oxidative and Non-Oxidative Bacterial Killing by Human Neutrophils

doi: 10.3389/fphar.2020.554353

Figure Lengend Snippet: Bacterial killing by neutrophil granule extracts and neutrophils with and without mucins. (A) Human neutrophil granule extracts markedly inhibited Pseudomonas aeruginosa (PAO1) growth but did not affect Burkholderia cenocepacia (C5424) density (n = 3). (B) Characterization of CF airway mucins (CFAM) and salivary mucins of healthy volunteers (SM). Samples of CFAM (200 μg) and SM (200 μg) were separated on a 0.7% agarose gel. The proteins were transferred to nitrocellulose membranes and revealed with either goat polyclonal mucin MUC5B IgG or MUC5AC concentrated Clone B442 antibody. (C) Neutrophil killing of B. cenocepacia (n = 9) and (D) P. aeruginosa (n = 9) in the absence or presence of CFAM or (E) SM (n = 7). CFAM and SM suppressed neutrophil-mediated killing of PAO1 and C5424 bacteria after incubation for 2 h at 37°C. (n = 9, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: The membranes were then incubated with goat polyclonal MUC5B IgG (1:250, Santa Cruz Biotechnology, Santa Cruz, CA) or MUC5AC-Concentrated-Clone B442 (1:250, Biomeda, Foster City, CA) overnight at 4°C and washed three times in TBS.

Techniques: Agarose Gel Electrophoresis, Incubation

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Structural Diversity of Human Gastric Mucin Glycans *

doi: 10.1074/mcp.M117.067983

Figure Lengend Snippet: In situ proximity ligation assay (PLA) showing human gastric MUC5AC decorated with ABH-blood group antigens. Primary antibodies recognizing MUC5AC and blood type A (A), B (B), and H (C) were used. The PLA signal was shown in red fluorescent spots indicating that MUC5AC and corresponding blood group antigens were in close proximity. The nuclei were stained blue with DAPI. The autofluoresence of tissue (green) delineated the tissue structure.

Article Snippet: Primary antibodies against blood type H (monoclonal mouse anti-human blood group H antigen, clone A70-A/A9, at a concentration of 2.5 μg/ml, ThermoFisher Scientific, Waltham, MA), A (monoclonal mouse anti-human blood group A, clone HE-193, dilution 1:80, ThermoFisher Scientific), B (monoclonal mouse anti-human blood group B, clone HEB-29, dilution 1:40, Abcam, Cambridge, UK), and MUC5AC (polyclonal rabbit anti-oligomeric mucus/gel-forming MUC5Ac N-term aa552–567, at a concentration of 5 μg/ml, antibodies-online GmbH, Aachen, Germany) were used and incubated at 4 °C overnight.

Techniques: In Situ, Proximity Ligation Assay, Staining