Overnight cultures of the wildtype strain E8202 and its mutant E8202 ptsD::GM were diluted 1:100 in BHI, BHI with 2 g/L mannose, Müller Hinton (MH) broth, MH with 2 g/L mannose (Sigma-Aldrich, US), Lysogeny broth (LB) or LB with 2 g/L mannose and growth was measured in an Epoch 2 Spectrophotometer with Gen5 Software (BioTek Instruments Inc., Winooski, Vermont US) at 37°C, shaking at 425 rpm, with OD600 measurement every 10th min for 18 h (n=6, biological triplicates, technical duplicates). E8202 and its mutant E8202 ptsD::GM were subjected to the spot on lawn assay as described above and in addition to BHI, the assay was also conducted on BHI with 2 g/L mannose, LB, and MH agar plates.
Mannose
Manufactured by Merck Group
Sourced in United States, Germany, China, Sao Tome and Principe, Italy, Japan, France, Macao, Sweden
Mannose is a type of sugar molecule that is commonly used in laboratory settings. It serves as a core structural component in various biological compounds and can be utilized in a variety of applications within the scientific research field.
Automatically generated - may contain errors
Lab products found in correlation
Galactose, by Merck Group (115 mentions)
Xylose, by Merck Group (107 mentions)
Arabinose, by Merck Group (91 mentions)
Rhamnose, by Merck Group (76 mentions)
Fucose, by Merck Group (60 mentions)
Fructose, by Merck Group (45 mentions)
Glucuronic acid, by Merck Group (39 mentions)
Galacturonic acid, by Merck Group (26 mentions)
Ribose, by Merck Group (22 mentions)
Sucrose, by Merck Group (18 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (17 mentions)
Lactose, by Merck Group (15 mentions)
Monosaccharide standard, by Merck Group (14 mentions)
Maltose, by Merck Group (13 mentions)
Glucosamine, by Merck Group (10 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (8 mentions)
Furfural, by Merck Group (8 mentions)
Acetic acid, by Merck Group (8 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (8 mentions)
Bovine serum albumin (bsa), by Merck Group (8 mentions)
182 protocols using mannose
1
Growth Profiling of E. coli Mutant
2
Quantifying Cellular Uptake of Mannose-IPC-SPIOs
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M2 macrophages were seeded into 24-well polystyrene TC-treated microplates. Some cells were incubated with 500 μg/mL of mannose (Sigma) for 2 hours to partially block mannose receptors and rinsed twice with PBS with calcium and magnesium. Cells were exposed to 150 μg/mL of mannose-IPC-SPIOs in cell culture media for 1, 3, or 6 hours. Cell media was aspirated, and cells were rinsed twice with PBS supplemented with calcium and magnesium. Control cells were not exposed to nanoparticles. The absorbance of nanoparticles attached to cells was measured at 450 nm using a plate reader (BioTek). Control cells were used to obtain baseline cellular absorbance, and all other data points were compared to this control sample.
3
Chitosan-based Vaccines Encapsulating KAg
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chitosan-based NPs encapsulating KAg (CS NPs-KAg) and mannosylated chitosan-based NPs encapsulating KAg (mCS NPs-KAg) vaccines were prepared using an ionic gelation method as previously described (24 (link)). For mannose-conjugated chitosan (mCS) preparation, 40 mg each of mannose (Sigma, MO) and sodium triacetoxyborohydride (Sigma, MO) mixture in 0.2 M borate buffer was slowly added into 200 mg chitosan [1% (w/v)] suspension (25 (link)) under magnetic stirring for 72 h at 56°C (34 (link)). The mCS was dialyzed against milli-Q-water for 48 h. Both chitosan and mannose modified chitosan were dissolved in 1% acetic acid solution. Twenty milligrams of mCS or chitosan were added into 20 ml milli-Q-water under magnetic stirring, pH was adjusted to 4.3 and mixed with 2 mg KAg (H1N2-OH10) in 3-(N-morpholino) propanesulfonic acid (MOPS) buffer pH 7.4. Followed by tripolyphosphate [1% (w/v) (Sigma, MO)] 5 mg in 10 ml milli-Q-water was added dropwise and the mCS NPs-KAg or CS NPs-KAg vaccines were obtained after centrifugation at 10,976 × g for 30 min, washed, dispersed in milli-Q-water and used for vaccination. Both the vaccines had ~80% antigen encapsulation efficiency and characterized as described previously (24 (link), 25 (link)). Both the chitosan-based vaccine formulations were freshly prepared and used in animals.
4
HUVEC Culture and Glucose Treatments
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Human umbilical vein endothelial cells (HUVECs) were obtained from ScienCell Research Laboratories and were cultured in endothelial cell medium supplemented with 1% penicillin-streptomycin solution and 5% foetal bovine serum in a humidified incubator containing 5% CO 2 at 37 °C. For the treatments, HUVECs were incubated with normal glucose (5.5 mM), high glucose (HG; 25 mM) or a high concentration of mannose (OS; 5.5 mM glucose and 19.5 mM mannose) for 48 h. mannose and glucose were purchased from Sigma-Aldrich (St. Louis, MO, USA).
5
Polymer-Based Antigen Delivery System
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We used Cs (#24601, HMC+, Germany); PK, PR, PE, and PD (#PLKC250, #PLR200, #PLE200, #PLD200, Alamanda Polymers, USA); Saccharides including sorbitol (#13273, Thermo Fisher Scientific, USA), glucose (#G7021, Merck, Germany), alginic acid (#A0733, TCI, Japan), mannose (#M6020, Merck, Germany), or mannans (#M7504, Merck, Germany); OVA (LS003059, Worthington Biochemical, USA); Alexa Fluor™ 660 Protein Labeling kit (#A20171, Thermo Fisher Scientific, USA); LPS (#L4391, Merck, Germany); and GM-CSF (#315–03, PeproTech, USA); DMSO (#D8418, Merck, Germany); HCl (#1943–0150, Showa, Japan); NaOH (#0812–0150, Showa, Japan).
6
Protein-Sugar Solution Preparation
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BSA, sucrose, and sucralose were procured from Sigma-Aldrich (St.
Louis, MO). Mannose and trehalose were purchased from Merck &
Co. Inc. (Kenilworth, NJ). BSA concentrations of ∼30, 40, 60,
80, and 90 mg mL–1 were prepared by dissolving appropriate
amounts of lyophilized BSA powder in milliQ water. For preparation
of protein–sugar solutions, 0.1, 0.4, and 0.7 M solutions of
trehalose, sucrose, and Mannose were prepared, to which BSA was added
to obtain constant protein concentrations of 40 mg mL–1. For sucralose, the highest concentration of sugar used was limited
to 0.5 M due to its low solubility in water. The protein–sugar
solutions were hence prepared with sucralose concentrations of 0.1,
0.3, and 0.5 M. Protein concentrations were spectrophotometrically
verified by UV spectroscopy at 280 nm using a Cary 50 Bio UV–vis
spectrometer (Agilent Technologies, Santa Clara, CA). UV spectra of
the prepared protein and protein–sugar solutions were recorded
using a 1 mm cuvette and an absorption coefficient of 42 925
M–1 cm–1.32
Louis, MO). Mannose and trehalose were purchased from Merck &
Co. Inc. (Kenilworth, NJ). BSA concentrations of ∼30, 40, 60,
80, and 90 mg mL–1 were prepared by dissolving appropriate
amounts of lyophilized BSA powder in milliQ water. For preparation
of protein–sugar solutions, 0.1, 0.4, and 0.7 M solutions of
trehalose, sucrose, and Mannose were prepared, to which BSA was added
to obtain constant protein concentrations of 40 mg mL–1. For sucralose, the highest concentration of sugar used was limited
to 0.5 M due to its low solubility in water. The protein–sugar
solutions were hence prepared with sucralose concentrations of 0.1,
0.3, and 0.5 M. Protein concentrations were spectrophotometrically
verified by UV spectroscopy at 280 nm using a Cary 50 Bio UV–vis
spectrometer (Agilent Technologies, Santa Clara, CA). UV spectra of
the prepared protein and protein–sugar solutions were recorded
using a 1 mm cuvette and an absorption coefficient of 42 925
M–1 cm–1.32
7
Bacterial Identification Protocol
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The test sample was a combined stock isolate from previous ARTIs research totaling 466 single bacterial isolates. This sample has been purified from bacterial contaminants, and rejuvenated.
The bacterial growth medium used was Mueller Hinton Agar (MHA) (Oxoid) with a concentration of 38 g/L, according to the guidelines from the CLSI 14 .
Biochemical test materials for bacterial identification include Lactose (Merck), Mannose (Merck), Maltose (Merck), peptone (Oxoid), phenol red (Taylor), Kovac`s reagent (Bio-Rad), TSIA , methyl red (HiMEdia), α-naphthosl (Merck).
The bacterial growth medium used was Mueller Hinton Agar (MHA) (Oxoid) with a concentration of 38 g/L, according to the guidelines from the CLSI 14 .
Biochemical test materials for bacterial identification include Lactose (Merck), Mannose (Merck), Maltose (Merck), peptone (Oxoid), phenol red (Taylor), Kovac`s reagent (Bio-Rad), TSIA , methyl red (HiMEdia), α-naphthosl (Merck).
8
Monosaccharide Composition Analysis by HPLC
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EPS monosaccharide composition was determined by acid hydrolysis of the lyophilized and rehydrated samples using 5% (v/v) perchloric acid (70%) at 100 °C for 5 h [42 (link)]. After filtration, hydrolyzed samples were analyzed using a HPLC Dionex DX500 system equipped with a GP50 gradient pump, an ED40 Electrochemical Detector in Pulsed Amperometric Detection (HPAEC-PAD), and Dionex PeakNet 5.11 chromatographic software. The chromatographic separation of sugars was carried out using a Dionex CarboPac PA1 column heated at 30 °C and a Carbopac PA1 guard column in isocratic mode with an elution of 150 mm NaOH at a flow rate of 1.0 mL/min [43 (link)]. Identification and quantification of the main monosaccharides was performed by integrating calibration peaks obtained from the relevant standards solutions of arabinose, glucose, fructose, and mannose, all purchased from Merck KGaA (Darmstadt, Germany).
9
Lignocellulosic Biomass Characterization
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All used chemicals were of analytical reagent grade. Acetic acid, dichloromethane, sulfuric acid (98%), sodium hydroxide, methanol, hydrochloric acid, suprapure nitric acid 65%, ethanol, toluene, acetone, hydrogen peroxide 30%, glucose, mannose, galactose, arabinose, pyridine, hydroxylamine hydrochloride (NH2OH·HCl) and salicin were purchased from Merck (Darmstadt, Germany). Sodium chlorite (80%) was purchased from Alfa Aesar GmbH & Co (Karlsruhe, Germany). Enzymes cellulase from Trichoderma reesei ATCC 26921, β-glucosidase from almonds, S. cerevisiae YSC2, peptone, BSTFA, 5-hydroxymethylfurfural and furfural were purchased from Sigma-Aldrich (St. Louis, MO, USA). Trichoderma reesei ATCC 26921 is a lyophilized powder with concentration ≥1.0 unit per mg of solid. All solutions were prepared by using ultrapure water (18.2 MΩcm−1 at 20 °C) obtained from a Direct-Q3 UV Water Purification System (Millipore, Molsheim, France).
10
Monosaccharide Profiling of Fucoidan by HPLC
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The content of monosaccharides in fucoidan was determined after acid hydrolysis. Five milligrams of fucoidan standard (Sigma-Aldrich. Co,. Spruce St. Louis, USA, Product of Australia) was dissolved in 1 ml HPLC grade water then treated with 1 ml of 4 M trifluoroacetic acid. The solution was heated for 2 hours at 100°C then filtered by 0.45 µm pore size filter (Merck KGaA, Darmstadt, Germany). The filtrate was neutralized with 0.1M NaOH and then evaporated with liquid nitrogen until the remaining volume of 0.5 ml. The content of monosaccharides of the fucoidan hydrolyzate was analyzed by High-Performance Liquid Chromatography (HPLC) (Shimadzu, SIL-10AD) using a refractive index detector (RID) equipped with a ligand-exchange column for Carbohydrate (Agilent Hi-Plex H, 7.7 × 100 mm) eluted with 0.005 M H 2 SO 4 at a flow rate of 0.7 ml/min. A sample of 20 µl was injected with separation time of 30 minutes. The standard monosaccharides used were fucose (Nacalai Tesque, Kyoto, Japan), glucose (Merck KGaA, Darmstadt, Germany), galactose (Merck KGaA, Darmstadt, Germany), rhamnose (Cica Reagen, Kanto Chemical, Tokyo, Japan), mannose (Merck KGaA, Darmstadt, Germany) and xylose (Merck KGaA, Darmstadt, Germany).
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