polyclonal rabbit polyclonal anti muc5ac antibody  (Boster Bio)

Journal: bioRxiv

Article Title: Requirement for Fucosyltransferase 2 in Allergic Airway Hyperreactivity and Mucus Obstruction

doi: 10.1101/2024.05.10.593580

Figure Lengend Snippet: A . α-Fuc was detected using the lectin Ulex europaeus agglutinin-1 (UEA1, yellow, 8 μg/ml), which labeled airway mucous cells and a mucus plug in a person who died from fatal asthma. The airway mucins MUC5AC (magenta, 1:1,000) and MUC5B (cyan, 1:500) were both detected. B-F . UEA1 labels mouse airway epithelium after induction of allergic inflammation using Aspergillus oryzae extract (AOE), and α-Fuc detection is Fut2-dependent. In C57BL/6 mice exposed to AOE ( B ), MUC5B (cyan, 1:500), MUC5AC (magenta, 1:2,500) and α-Fuc (UEA1, yellow, 8 μg/ml) labeling co-localize within airway epithelia. In Fut2 +/+ mice at baseline ( C ), α-Fuc and Muc5ac are not detectable in airway epithelia, but Muc5b is observed. After AOE, Muc5b is sustained while Muc5ac and α-Fuc detection are both induced in Fut2 +/+ animals ( D ). In Fut2 −/− mice, baseline and AOE-induced expression of Muc5b ( E ) and Muc5ac ( F ) are sustained, but AOE-induced α-Fuc detection is lost ( F ). Nuclei are stained with DAPI (blue). Scale bars: 100 μm for low magnification images and 50 μm for insets in A and C-F ; 10 μm for low magnification image and 8 μm for insets in B .

Article Snippet: Mouse mucins were detected using polyclonal rabbit-anti-mouse MUC5AC produced against the peptide N-CHALGDTSHAESSEQEFKSKESEEHGQQLAFR (Pacific Immunology, Ramona, CA, 1:2500 dilution), mouse-anti-mouse MUC5B (clone MDA-3E1, Cat. MABT899, MilliporeSigma, Burlington, MA, 1:500 dilution), and UEA1 (as above).

Techniques: Labeling, Expressing, Staining

Journal: bioRxiv

Article Title: Requirement for Fucosyltransferase 2 in Allergic Airway Hyperreactivity and Mucus Obstruction

doi: 10.1101/2024.05.10.593580

Figure Lengend Snippet: Lung lavage fluid was obtained from saline and AOE challenged Fut2 +/+ (black) and Fut2 −/− (magenta) mice. A , B . AOE challenge resulted in increases in total numbers of cells ( A ) and eosinophils ( B ), which did not differ between Fut2 +/+ and Fut2 −/− mice. Data were compared using groups by Kruskal-Wallis ANOVA. ‘*’, p < 0.05 using Dunn’s post-hoc test for multiple comparisons between genotype and challenge groups (p-values are shown in A and B ). Comparisons were made between AOE challenged Fut2 +/+ mice (n = 9 biological replicates), AOE challenged Fut2 −/− mice (magenta, n = 10 biological replicates), saline challenged Fut2 +/+ mice (n = 10 biological replicates), and saline challenged Fut2 −/− mice (magenta dashed lines, n = 9 biological replicates). Solid lines and filled circles identify AOE challenged animals; dashed lines and open circles depict saline challenged controls. C . Combined immuno- and lectin blotting was performed on neat lavage. Equal volumes of lavage fluid (27 μl) were loaded per well, separated in 1% SDS agarose gels, and transferred to PVDF membranes. Membranes were probed with biotinylated UEA1 (1:1,000, 2 μg/ml) and either rabbit-anti-MUC5AC (1:2,000) or rabbit-anti-MUC5B (1:5,000). For secondary detection, Alexa 680-conjugated streptavidin and Alexa 800-conjugated labeled goat-anti-rabbit probes (Licor, 1:15,000) were applied. Monochrome images were acquired and pseudocolored magenta (MUC5AC), cyan (MUC5B), or yellow (UEA1). Image overlays demonstrate UEA and mucin signal colocalization (white in merged panels).

Article Snippet: Mouse mucins were detected using polyclonal rabbit-anti-mouse MUC5AC produced against the peptide N-CHALGDTSHAESSEQEFKSKESEEHGQQLAFR (Pacific Immunology, Ramona, CA, 1:2500 dilution), mouse-anti-mouse MUC5B (clone MDA-3E1, Cat. MABT899, MilliporeSigma, Burlington, MA, 1:500 dilution), and UEA1 (as above).

Techniques: Saline, Labeling

Journal: Cell reports

Article Title: Long-Term In Vitro Expansion of Epithelial Stem Cells Enabled by Pharmacological Inhibition of PAK1-ROCK-Myosin II and TGF-β Signaling

doi: 10.1016/j.celrep.2018.09.072

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-MUC5AC , Santa Cruz Biotechnology , Cat# sc-20118; RRID: AB_2146854.

Techniques: Virus, Selection, Recombinant, Real-time Polymerase Chain Reaction, Sequencing, Paraffin Section, Staining, Enzyme-linked Immunosorbent Assay, In Vivo, Methylated DNA Immunoprecipitation Sequencing, Cell Culture, Software, DNA Methylation Assay

Journal: The Journal of pathology

Article Title: Hypoxia-inducible factor-1 signalling promotes goblet cell hyperplasia in airway epithelium

doi: 10.1002/path.2863

Figure Lengend Snippet: Hypoxia-inducible signalling is detected in areas of goblet cell hyperplasia in large airways of patients with COPD. (A) Representative images of bronchial epithelium from former smokers without COPD; top row, pseudostratified ciliated bronchial epithelium with normal ratio of basal, ciliated and goblet cells; bottom row, pseudostratified bronchial epithelium with hyperplasia of goblet cells. (B) Representative images of bronchial epithelium from patients with COPD; top row, normal-appearing pseudostratified ciliated bronchial epithelium with normal ratio of basal, ciliated and goblet cells; bottom row, pseudostratified bronchial epithelium with hyperplasia of goblet cells. There was a significant increase of goblet cell size and an approximate doubling of the height of the epithelial layer in COPD compared to former smokers without COPD. Column I, H & E-stained tissue sections; column II, PAS-stained tissue sections; column III, IHC with anti-MUC5AC antibody (brown stain); MUC5AC-positive granules were detected in single cells in normal-appearing pseudostratified epithelium (top) and in goblet cells with hyperplasia (bottom); column IV, IHC with anti-HIF-1α antibody. Cytoplasmic detection of HIF-1α was rarely observed in epithelial cells in former smokers without COPD and in normal-appearing bronchial epithelium in patients with COPD, while nuclear detection was observed in goblet cell hyperplasia in COPD. All images, × 400 magnification.

Article Snippet: To detect MUC5AC protein in apical washings, we used a rabbit polyclonal anti-MUC5AC antibody (clone H-160, No. 20 118; Santa Cruz Biotechnology) and HRP (horseradish peroxidase)-conjugated goat anti-rabbit polyclonal antibody (No. 31 462; Thermo Scientific, Rockford, IL, USA).

Techniques: Staining

Journal: The Journal of pathology

Article Title: Hypoxia-inducible factor-1 signalling promotes goblet cell hyperplasia in airway epithelium

doi: 10.1002/path.2863

Figure Lengend Snippet: Hypoxia and HIF-1α stabilization affect HBEC differentiation towards mucus-producing cell hyperplasia in vitro. (A) H & E-stained paraffin sections. Normal-appearing pseudostratified ciliated epithelium was noted in untreated control cultures compared to hyperplasia of mucus-producing cells in both hypoxic and BiPS-treated cultures. (B) IHC with anti-MUC5AC antibody (brown stain). Increased proportion of MUC5AC-positive cells situated between columnar surface cells in hypoxia- and BiPS-treated cultures. All images ×600 magnification. (C) Graph showing MUC5AC protein concentrations in apical washings. *p < 0.01 compared to untreated control (normoxia); **p < 0.01 compared to vehicle-treated control.

Article Snippet: To detect MUC5AC protein in apical washings, we used a rabbit polyclonal anti-MUC5AC antibody (clone H-160, No. 20 118; Santa Cruz Biotechnology) and HRP (horseradish peroxidase)-conjugated goat anti-rabbit polyclonal antibody (No. 31 462; Thermo Scientific, Rockford, IL, USA).

Techniques: In Vitro, Staining, Control