Muc5b

Manufactured by Santa Cruz Biotechnology

Sourced in United States

MUC5B is a laboratory product offered by Santa Cruz Biotechnology. It is a protein that can be used for research purposes. No further details on the core function or intended use of this product can be provided in an unbiased and factual manner.

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Lab products found in correlation

Muc5ac, by Santa Cruz Biotechnology (3 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions) Cy3 secondary antibody, by Jackson ImmunoResearch (1 mentions) Foxa2, by Merck Group (1 mentions) Muc5ac, by Novus Biologicals (1 mentions) Normal goat serum (ngs), by Jackson ImmunoResearch (1 mentions) Tx 100, by Thermo Fisher Scientific (1 mentions) Alexa fluor 555 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Phalloidin 633, by Thermo Fisher Scientific (1 mentions) Goat anti mouse alexa fluor 488 and, by Thermo Fisher Scientific (1 mentions) Superblock, by Thermo Fisher Scientific (1 mentions) Cellsens, by Olympus (1 mentions) Fluoview fv3000 confocal microscope, by Olympus (1 mentions) Fluorescently labeled secondary antibody, by Thermo Fisher Scientific (1 mentions) Sc 20119, by Santa Cruz Biotechnology (1 mentions) Vs200 slide scanner, by Olympus (1 mentions)

5 protocols using muc5b

Quantifying Muc5ac, Muc5b, IL-6, and KC

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We measured Muc5ac and Muc5b (Santa Cruz, USA) protein levels with ELISA, as previously described.12 (link),26 (link) The concentration of IL-6 (eBioscience, USA) and keratinocyte-derived cytokine (KC) (R&D Systems Inc.,USA) were determined with ELISA kits following the manufacturers’ protocol.

Song Y., Wang W., Xie Y., Xiang B., Huang X., Guan W, & Zheng J. (2019). Carbocisteine inhibits the expression of Muc5b in COPD mouse model. Drug Design, Development and Therapy, 13, 3259-3268.

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Immunofluorescence Analysis of Respiratory Syncytial Virus

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For immunofluorescence (IF) analysis, HBECs were plated in the 24‐well plate with cover glass, fixed with 4% paraformaldehyde permeabilized with 0.1% Triton X‐100 in PBS for 15 minutes at room temperature. After blocking with 1% BSA, the HBECs were incubated with primary antibodies to RSV‐F (Abcam, ab24011), MUC5AC (Santa Cruz, sc‐21701) or MUC5B (Santa Cruz, sc‐21768) overnight at 4°C. Subsequently, Cy3 secondary antibody (Jackson ImmunoResearch, 715‐165‐150) was used to probe the primary Ab. DAPI was used for nuclear staining.²⁷ IF samples were visualized using a fluorescence microscope (Carl Zeiss MicroImaging GmbH, Göttingen, Germany). Images were captured with a digital camera (Axio‐Cam ICc3, Spectra Service, Ontario, NY, USA) and analysed with AxioVision Rel. 4.7 software (Zeiss).

Du X., Yang Y., Xiao G., Yang M., Yuan L., Qin L., He R., Wang L., Wu M., Wu S., Feng J., Xiang Y., Qu X., Liu H., Qin X, & Liu C. (2020). Respiratory syncytial virus infection‐induced mucus secretion by down‐regulation of miR‐34b/c‐5p expression in airway epithelial cells. Journal of Cellular and Molecular Medicine, 24(21), 12694-12705.

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Quantifying Nasal Epithelial Cell Types

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Human nasal tissues were stained with PAS, and the PAS-positive cells, which displayed a purple color, were counted via light microscopy (400×) and expressed as the percentage of the total epithelial cells (500 cells counted). IHC was performed as reported elsewhere,13 (link) and the sections were stained using mouse monoclonal antibodies against the following proteins: FoxA2 (1:50; Millipore, Billerica, MA, USA), MUC5AC, and MUC5B (1:100; Santa Cruz Biotech, Santa Cruz, CA, USA). Then, the antibodies were detected via streptavidin-biotin-horseradish peroxidase complex formation. The immunostaining result was considered positive when brown cells appeared following reaction with the reagent 3', 3'-diaminobenzidine. Replacement of primary antibodies with isotype-matched IgG was used as a negative control. The sections were examined via light microscopy (400×), and the patterns of antibody staining were scored in a quantitative manner. The pattern of immunoreactivity was analyzed in the epithelium and subepithelial area. The number of immunoreactive cells (stained brown) was expressed as the percentage of the total cell number (500 cells counted). The immunoreactivity score was assessed by 2 independent observers in a blinded manner, and their results were averaged.

Luo Q., Zhang J., Wang H., Chen F., Luo X., Miao B., Wu X., Ma R., Luo X., Xu G., Shi J, & Li H. (2015). Expression and Regulation of Transcription Factor FoxA2 in Chronic Rhinosinusitis With and Without Nasal Polyps. Allergy, Asthma & Immunology Research, 7(5), 458-466.

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Immunocytochemistry of Newborn Piglet Trachea

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Trachea were excised from newborn piglets and immediately fixed in 4% paraformaldehyde (EMS) in PBS for 1 hr at room temperature. Tissues were then placed in 30% sucrose and incubated overnight at 4°C, followed by quick-freezing in OCT using a dry ice/EtOH bath and stored at −80°C. Prior to immunocytochemistry, frozen blocks of tissue were cryosectioned at 7 μm followed by permeabilization in 0.3% TX-100 (Thermo-Fisher) in PBS for 20 min, and blocked in Super-Block (Thermo-Fisher) with 5% normal goat serum (Jackson ImmunoResearch) for 1 hr, all at room temperature. Tissue sections were then incubated for 2 hr at 37°C with indicated antibodies: β-tubulin IV(1:300, Biogenex), MUC5AC (1:5000, Novus Biologicals), MUC5B (1:2000, Santa Cruz). Sections were then incubated for 1 hr with secondary antibodies goat-anti-mouse Alexa-Fluor-488 and goat anti-rabbit Alexa-Fluor-555 (1:1000, Molecular Probes/Invitrogen) and phalloidin-633 (1:300, Molecular Probes/Invitrogen). Slides were imaged on an Olympus Fluoview FV3000 confocal microscope with a Plan.ApoN 60X oil lens. Images were post-processed using the Olympus imaging software, CellSens.

Ostedgaard L.S., Price M.P., Whitworth K.M., Abou Alaiwa M.H., Fischer A.J., Warrier A., Samuel M., Spate L.D., Allen P.D., Hilkin B.M., Romano Ibarra G.S., Ortiz Bezara M.E., Goodell B.J., Mather S.E., Powers L.S., Stroik M.R., Gansemer N.D., Hippee C.E., Zarei K., Goeken J.A., Businga T.R., Hoffman E.A., Meyerholz D.K., Prather R.S., Stoltz D.A, & Welsh M.J. (2020). Lack of airway submucosal glands impairs respiratory host defenses. eLife, 9, e59653.

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Histological Analysis of HBEC Cultures

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HBEC grown on transwell inserts were fixed in 10% neutral buffered formalin for one hour, washed in DPBS, and stored until embedding in 70% ethanol at 4°C. Inserts were paraffin-embedded sectioned, and Hematoxylin and eosin (H&E) and AB-PAS stained by the UNC Pathology Services Core. Immunohistochemistry (IHC) was performed as previously described^{29 (link)}. Antibodies used for IHC were MUC5AC (Invitrogen MA5-12178) and MUC5B (Santa-Cruz sc-20119) each at 1:1000 dilution. Fluorescently-labeled secondary antibodies (Invitrogen) were used for mouse (A32773) and rabbit (A32795) at 1:1000 dilutions. Image acquisition was performed using a VS200 slide scanner (Olympus).

Penna T.C., Mazzola P.G, & Silva Martins A.M. (2001). The efficacy of chemical agents in cleaning and disinfection programs. BMC Infectious Diseases, 1, 16.

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