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Hog gastric mucin

Manufactured by Merck Group
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Sourced in United States, Germany, China

Hog gastric mucin is a purified glycoprotein derived from the gastric mucosa of pigs. It is used as a laboratory reagent and research tool to study various biological processes, such as cell signaling, protein-protein interactions, and mucosal function.

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Lab products found in correlation

D glucose, by Merck Group (2 mentions) Tryptone, by Thermo Fisher Scientific (2 mentions) L threonine, by Merck Group (2 mentions) Peptone, by Thermo Fisher Scientific (1 mentions) Bacto casitone, by BD (1 mentions) Escherichia coli atcc 25922, by American Type Culture Collection (1 mentions) Difco lactobacilli mrs broth, by BD (1 mentions) N acetylglucosamine, by Merck Group (1 mentions) L fucose, by Merck Group (1 mentions) N acetylgalactosamine, by Merck Group (1 mentions) Balb c mice, by Charles River Laboratories (1 mentions) Yeast extract, by Merck Group (1 mentions) Brain heart infusion medium, by Hopebio (1 mentions)

Spelling variants of this product

Mucin (95 citations) Hog gastric mucin type III (7 citations) Gastric mucin (5 citations)

7 protocols using hog gastric mucin

1

Cultivation of Akkermansia muciniphila

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Akkermansia muciniphila MucT (ATTC BAA-835) was grown in a basal medium as described previously, except without the addition of rumen fluid (Derrien et al., 2004 (link)). The medium was supplemented with either hog gastric mucin (0.5 %, Type III; Sigma-Aldrich, St. Louis, MO, USA) purified by ethanol precipitation as described previously (Miller and Hoskins, 1981 (link)), or D-glucose (10 mM, Sigma–Aldrich). The medium with glucose was also supplemented with BactoTM casitone (BD, Sparks, MD, USA), BBLTM yeast extract (BD), tryptone (Oxoid Ltd, Basingstoke, Hampshire, England), peptone (Oxoid ltd) (2 g/l each) and L-threonine (Sigma–Aldrich) (2 mM). Incubations were performed in serum bottles sealed with butyl-rubber stoppers at 37°C under anaerobic conditions provided by a gas phase of 182 kPa (1.5 atm) N2/CO2. Growth was measured by following optical density at 600 nm (OD600) using a spectrophotometer.
Ottman N., Huuskonen L., Reunanen J., Boeren S., Klievink J., Smidt H., Belzer C, & de Vos W.M. (2016). Characterization of Outer Membrane Proteome of Akkermansia muciniphila Reveals Sets of Novel Proteins Exposed to the Human Intestine. Frontiers in Microbiology, 7, 1157.
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2

In Vivo Antibacterial Efficacy Evaluation

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Escherichia coli ATCC 25922 was obtained from American Type Culture Collection (Manassas, VA, United States). Cells were grown overnight on solid agar, suspended in saline and adjusted to an OD of 0.1 at 625 nm. The suspension was further diluted in 5% hog gastric mucin (Sigma-Aldrich Corporation, St. Louis, MO, United States) and final inoculums between 4.8 × 105 and 2.0 × 106 colony forming units (cfu) per mouse were injected ip (500 μL). Api137 and levofloxacin was dissolved in sterile saline and sterile water, respectively, and were administered iv (5 mL/kg) into the tail vein or sc (10 mL/kg) into the back. Alternatively, an hour post infection ALZET® osmotic pumps were implanted under isoflurane anesthesia (see above). After 24 h, pumps were exchanged once and finally removed after 48 h. Animals were observed for 7 days post infection.
Knappe D., Schmidt R., Adermann K, & Hoffmann R. (2019). Continuous Subcutaneous Delivery of Proline-Rich Antimicrobial Peptide Api137 Provides Superior Efficacy to Intravenous Administration in a Mouse Infection Model. Frontiers in Microbiology, 10, 2283.
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3

Determining Lethal Dose and Bacterial Colonization in Mice

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Six-week-old female BALB/c mice were purchased from Dashuo Biotechnology Co., Ltd. (Chengdu, China). To determine the 50% lethality dose (LD50), bacteria were grown to OD600 of 0.8 to 0.9 and harvested by centrifugation at 3,452 × g at room temperature. The centrifuged S. Typhimurium, S. Enteritidis or their derivatives were resuspended and adjusted to the appropriate OD600 value by buffered saline with gelatin (BSG) [40 (link)]. In contrast, the centrifuged S. Paratyphi A or its derivatives were resuspended and adjusted to the appropriate OD600 value by 10% hog gastric mucin (Sigma). Six mice per group were infected orally with 20 μl of BSG or intraperitoneally injected with 500 μl 10% hog gastric mucin containing various doses of bacteria, ranging from 1 × 104 CFU to 1 × 108 CFU. Mice were monitored for mortality or signs of significant morbidity daily. The LD50 was calculated using the method of Reed and Muench. To evaluate colonization, three mice per group were orally inoculated with 20 μl of BSG containing 1 × 109 CFU bacteria. On days 4 and 8 post-inoculation, Peyer’s patches, spleen and liver samples were collected. Samples were homogenized, dilutions were plated onto MacConkey and LB agar to determine viable counts.
Li P., Zhang K., Lei T., Zhou Z, & Luo H. (2022). Multiple immunodominant O-epitopes co-expression in live attenuated Salmonella serovars induce cross-protective immune responses against S. Paratyphi A, S. Typhimurium and S. Enteritidis. PLOS Neglected Tropical Diseases, 16(10), e0010866.
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4

Cultivation of Gut Bacterial Species

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Akkermansia muciniphila MucT (ATTC BAA-835) was grown in a basal medium as described previously [7 (link)]. The medium was supplemented with either hog gastric mucin (0.5%, Type III; Sigma-Aldrich, St. Louis, MO, USA), a mix of sugars (D-glucose, L-fucose, N-acetylglucosamine, N-acetylgalactosamine; 2.5 mM each, Sigma-Aldrich) or glucose (10 mM, Sigma-Aldrich). The medium without mucin was supplemented with tryptone (8 g/l, Oxoid Ltd, Basingstoke, Hampshire, England) and L-threonine (2 mM, Sigma-Aldrich). Incubations were performed in serum bottles sealed with butyl-rubber stoppers at 37°C under anaerobic conditions provided by a gas phase of 182 kPa (1.5 atm) N2/CO2 (80/20 ratio). Growth was measured by spectrophotometer as optical density at 600 nm (OD600).
Faecalibacterium prausnitzii A2-165 was grown anaerobically at 37°C in YCFA medium supplemented with 33 mM glucose [23 (link)]. Lactobacillus plantarum WCFS1 was grown aerobically and Bifidobacterium breve DSM-20213 anaerobically at 37°C in Difco™ Lactobacilli MRS broth (Becton Dickinson, Sparks, USA).
Ottman N., Reunanen J., Meijerink M., Pietilä T.E., Kainulainen V., Klievink J., Huuskonen L., Aalvink S., Skurnik M., Boeren S., Satokari R., Mercenier A., Palva A., Smidt H., de Vos W.M, & Belzer C. (2017). Pili-like proteins of Akkermansia muciniphila modulate host immune responses and gut barrier function. PLoS ONE, 12(3), e0173004.
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5

Passive Transfer of Antibodies Protects Mice Against Paratyphoid

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All animal studies were performed in facilities accredited by the Association for Assessment and Accreditation of Laboratory Animal Care and in compliance with policies established by the US Department of Agriculture Animal Welfare Act and Public Health Service. These animal experiments were conducted with approval from the Institutional Animal Care and Use Committee at the University of Maryland, School of Medicine. Pre-immunization and day 56 post-immunization rabbit sera were heat-inactivated at 56 °C for 30 min and diluted 1:10 in PBS. Naïve 6–8 week old female BALB/c mice (Charles River Laboratories, MA, USA) were injected i.p. with 100 μL of the diluted rabbit sera or PBS. After 5–6 hours, recipient mice were challenged i.p. with 7.3 × 103 CFU of S. Paratyphi A strain ATCC 9150 (prepared from an overnight HS plate) diluted in 500 μL of sterile 10% hog-gastric mucin (Sigma-Aldrich). Infected mice were monitored twice a day for three days, and animals that were moribund and/or had sustained 20% weight loss were euthanized and recorded as having succumbed to challenge. Vaccine efficacy for passively immunized mice was determined based on the proportionate attack rate (AR) in vaccinated and PBS control mice: ((ARcontrol – ARvaccinated)/ARcontrol) × 100.
Dhara D., Baliban S.M., Huo C.X., Rashidijahanabad Z., Sears K.T., Nick S.T., Misra A.K., Tennant S.M, & Huang X. (2020). Syntheses of Salmonella Paratyphi A associated oligosaccharide antigens and development towards anti-paratyphoid fever vaccines. Chemistry (Weinheim an der Bergstrasse, Germany), 26(68), 15953-15968.
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6

Canine Artificial Saliva Preparation

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Preparation of canine artificial saliva (CAS) was performed as described by Sanguansermsri et al. (2018) [8 (link)]. CAS was composed (per liter) by 1 g Lab Lemco Powder (Thermo Fisher Scientific, Denmark), 2 g yeast extract (Sigma-Aldrich, USA), 5 g proteose peptone (Merk, Germany), 2.5 g hog gastric mucin (Sigma-Aldrich, USA), 2.34 g NaCl, 1.5 g KCl, 0.1 g CaCl2 and 1.25 mL of 40% urea. The solution was sterilized by autoclave, except for urea that was filtered using a 0.22 µm cellulose acetate membrane filter and then added to the remaining components.
Cunha E., Rebelo S., Carneiro C., Tavares L., Carreira L.M, & Oliveira M. (2020). A polymicrobial biofilm model for testing the antimicrobial potential of a nisin-biogel for canine periodontal disease control. BMC Veterinary Research, 16, 469.
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7

Cultivation of Akkermansia muciniphila

Cited 1 time
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A. muciniphila MucT (ATCC BAA-835) was obtained from the American Type Culture Collection (ATCC). The strain was cultured as described previously [21 (link)]. The bacterial culture medium was brain–heart infusion medium (Qingdao Hope Bio-Technology Company, Qingdao, China) supplemented with hog gastric mucin (0.25% w/v Type II; Sigma-Aldrich, St. Louis, MO, USA). The bacterial cells were grown under anaerobic conditions at 37 °C for 18 h, and were enriched by centrifugation (6000× g, 10 min) and washed with normal saline. Finally, the bacterial sludge was resuspended in normal saline at the required final concentration.
Qi Y., Yu L., Tian F., Zhao J., Zhang H., Chen W, & Zhai Q. (2022). A. muciniphila Supplementation in Mice during Pregnancy and Lactation Affects the Maternal Intestinal Microenvironment. Nutrients, 14(2), 390.
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