Mannan from saccharomyces cerevisiae

Manufactured by Merck Group

Sourced in United States

Mannan from Saccharomyces cerevisiae is a laboratory product derived from the cell wall of the yeast Saccharomyces cerevisiae, commonly known as baker's yeast. It is a polysaccharide composed of mannose units. Mannan is used as a research tool for various applications in the life sciences.

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Lab products found in correlation

Resiquimod r 848, by Bio-Techne (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Elisa plate, by Corning (1 mentions) O phenylenediamine, by Merck Group (1 mentions) Chitosan, by Merck Group (1 mentions) Microtiter plate, by Thermo Fisher Scientific (1 mentions) 3 3 5 5 tetramethylbenzidine (tmb), by Thermo Fisher Scientific (1 mentions) Microplate reader, by Bio-Rad (1 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Dmem f12, by Corning (1 mentions) Dimethyl sulfoxide d6, by Cambridge Isotopes (1 mentions) Anti cd16 32 trustain fcx, by BioLegend (1 mentions) Fluorescein isothiocyanate isomer 1 fitc, by Merck Group (1 mentions) Barnstead nanopure system, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle s medium dmem with 4.5 g l glucose and l glutamine, by Corning (1 mentions) Raw264.7 macrophage, by American Type Culture Collection (1 mentions) Pen strep, by Thermo Fisher Scientific (1 mentions) Polyvinyl alcohol (pva), by Merck Group (1 mentions) Pe anti human cd209 dc sign antibody, by BioLegend (1 mentions) 30 μm pore filters, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Saccharomyces cerevisiae mannan Saccharomyces cerevisiae Mannan

21 protocols using mannan from saccharomyces cerevisiae

ELISA Assay for Complement C3 Activation

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ELISA plates (Costar) were coated with 50 μl of coating buffer (35 mM
Na₂CO₃, 15 mM NaHCO₃, pH 9.6) containing 100
μg/ml of mannan from Saccharomyces cerevisiae (Sigma) or
1% BSA (negative control) and incubated overnight at 4°C. After a one hour
blockage of the wells with blocking buffer (1% BSA in PBS), 1% NHS diluted
in GVB²⁺ was added together with different concentrations of
An. albimanus SGH or recombinant protein dissolved in PBS (final volume
of 100 μl/well) and incubated for 30 minutes at 37°C. Wells incubated with
serum and no inhibitor were used as positive controls. The wells were washed with PBS-Tw
(0.05% Tween-20 in PBS) and incubated for 60 minutes at room temperature with
blocking buffer containing anti-C3 antibody diluted 1:1000. After two washes, 50
μl of blocking buffer containing conjugated antibody (Sigma) diluted 1:1500 were
added to the wells. After two more washes, the wells were filled with 200 μL of
developing buffer (50 mM Na₃C₆H₅O₇, 50 mM
Na₂HPO₄, 1 mg/ml o-phenylenediamine (Sigma) and 0.075 %
H₂O₂, pH 5.0) and read at 450 nm and 37°C for 10 min in
the kinetic mode. The assays were done in duplicate and the means of the negative control
were subtracted of the other means. The results were transformed in percentage of C3
activation, considering the positive control as 100% of activation and then
analyzed using ANOVA and the Tukey test.

Mendes-Sousa A.F., Queiroz D.C., Vale V.F., Ribeiro J.M., Valenzuela J.G., Gontijo N.F, & Andersen J.F. (2016). An inhibitor of the alternative pathway of complement in saliva of new-world anopheline mosquitoes. Journal of immunology (Baltimore, Md. : 1950), 197(2), 599-610.

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Intranasal Mannan for Asthma Therapy

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Mannan from Saccharomyces cerevisiae was purchased from Sigma-Aldrich (St. Louis, MO) and prepared for Patented Use for Asthma Therapeutic [20 ]. Endotoxin level was <2 EU/mL. Mice that were in groups to receive MN treatment were given 1 mg MN in 10 μL intranasally one hour prior to each fungal antigen exposure.

Lew D.B., LeMessurier K.S., Palipane M., Lin Y, & Samarasinghe A.E. (2018). Saccharomyces cerevisiae-Derived Mannan Does Not Alter Immune Responses to Aspergillus Allergens. BioMed Research International, 2018, 3298378.

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Glycan-Based Immune Activation in BM-DCs

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After 4 days of derivation, the BM-DCs were stimulated for 1, 3 and 6 h with the following (n = 3 wells/treatment): RPMI-1640, 2.5 doses/well (equivalent to at least 2.5 × 10^3.5 EID₅₀) UV-inactivated IBV (Nobilis IB Ma5 Vet vaccine, MSD Animal Health), 100 µg/mL pustulan from Lasallia pustulata (InvivoGen, San Diego, CA, USA), 500 µg/mL Mannan from Saccharomyces cerevisiae (Sigma-Aldrich), 200 µg/mL chitosan (Sigma-Aldrich, cat. no. C3646), 10 µg/mL furfurman from Malassezia furfur (InvivoGen, cat. no. tlrl-ffm), 50 µg/mL MMG, or 50 µg/mL TDB. MMG and TDB were provided by Statens Serum Institut. Post stimulation, the BM-DCs were washed 3 times in ice-cold PBS, incubated in 10 mM EDTA on ice for 10 min and detached by thoroughly pipetting up and down. Subsequently, the cells were analysed by flow cytometry. The level of endotoxin contamination was determined by providers or previous studies to be insignificant in the glycan-based ligand concentrations used [34 (link),37 (link),38 (link)].

Larsen F.T., Guldbrandtsen B., Christensen D., Pitcovski J., Kjærup R.B, & Dalgaard T.S. (2020). Pustulan Activates Chicken Bone Marrow-Derived Dendritic Cells In Vitro and Promotes Ex Vivo CD4+ T Cell Recall Response to Infectious Bronchitis Virus. Vaccines, 8(2), 226.

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Complement Pathway ELISA Protocol

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Complement ELISAs were performed as previously described (37 (link)) with minor modifications. Briefly, microtiter plates (Nunc maxisorp) were pre-coated with 3 µg/mL IgM (Sigma) for classical pathway (CP) ELISA, with 20 µg/ml LPS (from Salmonella enteriditis, Sigma) for alternative pathway (AP) ELISA and with 20 µg/mL mannan (from Saccharomyces cerevisiae, Sigma) for lectin pathway (LP) ELISA. All coatings were prepared in 0.1 M carbonate buffer (pH 9) and incubated overnight at room temp. Plasma samples were diluted in Veronal Buffered Saline (VBS) + 0.1% gelatin + 5 mM MgCl₂ + 10 mM EGTA for AP ELISA and in VBS + 0.1% gelatin + 0.5 mM CaCl₂ + 0.25 mM MgCl₂ for CP ELISA and LP ELISA. After washing the plates with PBS 0.05% Tween-20, wells were blocked with 4% BSA in PBS-Tween. Then, plasma samples were added for 1 hour at 37°C in the appropriate buffer and dilutions. After additional washing, deposited C3b was detected using DIG-labelled mouse anti C3 antibody (WM1, 0.1 µg/mL) followed by Peroxidase conjugated sheep anti-DIG-Fab fragments (1:8000, Roche 11207733910). Finally, the plates were washed and developed using 3,3’,5,5’-tetramethylbenzidine (Thermo Fisher). The reaction was stopped by addition of 1 N H₂SO₄. Absorption at 450nm was measured using a microplate reader (Biorad).

de Vor L., Beudeker C.R., Flier A., Scheepmaker L.M., Aerts P.C., Vijlbrief D.C., Bekker M.N., Beurskens F.J., van Kessel K.P., de Haas C.J., Rooijakkers S.H, & van der Flier M. (2022). Monoclonal antibodies effectively potentiate complement activation and phagocytosis of Staphylococcus epidermidis in neonatal human plasma. Frontiers in Immunology, 13, 933251.

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Mannan-Targeted Polymeric Nanocarriers

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Poly (D,L-lactide-co-glycolide) (PLG) (50:50, ester terminated, M_W 7,000–17,000), Poly (vinyl alcohol) (PVA) (M_W 13,000–23,000, 87–89% hydrolyzed), dichloromethane (DCM), coumarin 6, fluorescein isothiocyanate isomer 1 (FITC) and mannan from Saccharomyces cerevisiae were purchased from Sigma (St. Louis, MO). α-D-mannopyranosyl 4-phenylisothiocyanate (MITC) was purchased from Synthose (Concord, ON, Canada). Ethanol was purchased from Decon Laboratories (King of Prussia, PA). Dimethyl sulfoxide (DMSO), annexin v binding buffer, fetal bovine serum (FBS), trypsin with 0.25% EDTA and pen/strep were purchased from Fisher (Hampton, NH). Ultrapure water was obtained from a Thermo Scientific Barnstead Nanopure system. Dimethyl sulfoxide d6 was purchased from Cambridge Isotope Labs (Tewksbury, MA). Concanavalin A rhodamine (ConA) was purchased from Vector Labs (Burlingame, CA). RAW 264.7 macrophages were obtained from ATCC (TIB-71). Dulbecco’s Modified Eagle’s Medium (DMEM) with 4.5 g/L glucose and L-glutamine and DMEM/F12 was purchased from Corning Cellgro (Corning, NY). Recombinant macrophage colony stimulating factor (MCSF) was purchased from Cell Guidance Systems (Cambridge, UK). Antibodies for flow cytometry, Trustain fcX (anti-CD16/32) and APC anti-mouse F4/80 antibody (clone BM8) were purchased from BioLegend (San Diego, CA, USA).

Isely C., Atube K.J., Cheung C.V., Steege C.F., Pellechia P.J, & Gower R.M. (2022). Surface Functionalization of Polymer Particles for Cell Targeting by Modifying Emulsifier Chemistry. ACS applied polymer materials, 4(4), 2269-2282.

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Inhibiting C-type Lectin Receptors with Mannan

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The antibodies used in the experiments were Mouse IgG2a Isotype Control (Invitrogen), MERS Coronavirus Spike Protein neutralizing Antibody MA5-29975 (Invitrogen), Invitrogen CD26 Monoclonal Antibody (BA5b) (Invitrogen), Human DC-SIGN+DC-SIGNR Antibody (R&D Systems), PE anti-human CD209 (DC-SIGN) Antibody (Biolegend), and CD26 Monoclonal Antibody (BA5b) PE (Invitrogen). As a lectin-binding control, mannan from Saccharomyces cerevisiae (Sigma-Aldrich) was used. Mannan is used in experiments to inhibit C-type lectin receptors, such as DC-SIGN due to its ability to block the interaction between these receptors and viral or pathogenic ligands. Being a complex polysaccharide, mannan competes with the binding sites of the receptors, acting as a decoy and preventing the attachment of infectious agents to host cells (Jan et al., 2018 (link)).

Labiod N., Luczkowiak J., Tapia M.M., Lasala F, & Delgado R. (2023). The role of DC-SIGN as a trans-receptor in infection by MERS-CoV. Frontiers in Cellular and Infection Microbiology, 13, 1177270.

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Glycoside Affinity Chromatography of Bacterial Pathogens

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Mannose-C-glycoside and 2FL-C-glycoside ketohydrazide resins, as prepared above in Part 1

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Stoppered spin columns with 30 μm pore filters (Thermo Scientific cat. 69725)

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E. coli ATCC 29425 (K12)

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C. jejuni NCTC 11168 (Cj11168)

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Brain-Heart Infusion broth and agar plates (BHI; BD cat. 237500)

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Mueller Hinton broth (BD cat. 275710)

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Citrated bovine blood (Quad Five cat. 930)

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Phosphate buffered saline (PBS)

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Mannose

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Galactose

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Mannan from Saccharomyces cerevisiae (Sigma cat. M7504)

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Lactose

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2′Fucosyllactose

Van Tassell M.L., Price N.P, & Miller M.J. (2014). Glycan-specific whole cell affinity chromatography: A versatile microbial adhesion platform. MethodsX, 1, 244-250.

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Antibody Labeling and Mannan Detection

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The primary antibodies used were mouse anti-dsRNA (J2 clone, Scicons), mouse pan-flavivirus (4G2 clone, Novus Biologicals), rabbit anti-MX1 (Thermo Fischer Scientific), mouse APC conjugated anti-langerin (clone 10E2, BioLegend), rabbit anti-langerin (clone D9H7R, Cell Signaling), recombinant anti-DC-SIGN (clone REA617, Miltenyi Biotec), mouse anti-CD1a (Novus Biologicals), FITC-conjugated anti-CD1a (clone HI149, Miltenyi Biotec), FITC-conjugated anti-HLA-DR (clone AC122, Miltenyi Biotec), mouse anti-HSP90 (clone F-8, Santa Cruz), and mouse anti-GAPDH (clone 6C5, Merck Millipore). Secondary antibodies were goat anti-mouse AF488 (Thermo Fisher Scientific), donkey anti-mouse AF568 (Thermo Fisher Scientific), donkey anti-rabbit AF647 (Thermo Fisher Scientific) and goat anti-mouse or anti-rabbit HRP conjugates (GE Healthcare).
Mannan from Saccharomyces cerevisiae was purchased from Sigma-Aldrich.

Martin M.F., Maarifi G., Abiven H., Seffals M., Mouchet N., Beck C., Bodet C., Lévèque N., Arhel N.J., Blanchet F.P., Simonin Y, & Nisole S. (2022). Usutu Virus escapes langerin-induced restriction to productively infect human Langerhans cells, unlike West Nile virus. Emerging Microbes & Infections, 11(1), 761-774.

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Peptide Inhibition of Cell Viability

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For the competition assays, P-113 and P-113Tri (12 μg/mL) were preincubated with 1, 2, 4, and 8 mg/mL mannan and laminarin at 4 °C for 30 min, followed by mixing with cells and incubating at 37 °C for 1 h. Then, the numbers of viable cells after peptide treatment were normalized to those of the control cells (no peptide treatment) and are reported as percentages. All carbohydrate stock solutions were prepared in 12.5 mM sodium acetate. Mannan from Saccharomyces cerevisiae and Laminarin from Laminaria digitata were purchased from Sigma-Aldrich (catalog numbers M7504 and L9634).

Lin G.Y., Chang C.F, & Lan C.Y. (2020). The interaction between Carbohydrates and the Antimicrobial Peptide P-113Tri is Involved in the Killing of Candida albicans. Microorganisms, 8(2), 299.

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Intratumoral Immunotherapy for Cancer

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Mannan from Saccharomyces cerevisiae, lipoteichoic acid (LTA) from Bacillus subtilis, and polyinosinic:polycytidylic acid (poly(I:C)) was obtained from Sigma-Aldrich (Saint Lous, MO, USA). BAM was obtained from NOF Corporation (White Plains, NY, USA). Resiquimod (R-848) was obtained from Tocris Bioscience (Minneapolis, MN, USA). Monoclonal anti-CD40 (clone FGK4.5/FGK45) was obtained from BioXCell (West Lebanon, NH, USA). Mannan-BAM synthesis was performed as previous reported (25 (link)). After development of tumors (average tumor volume 55 mm³), mice were treated intratumorally in days 0, 1, 2, 8, 9, 10, 16, 17, 18, 24, 25, and 26 with 50 μL of the therapeutic mixture consisting of 0.5 mg R-848 (HCl form), 0.5 mg poly(I:C), 0.5 mg LTA, and 0.4 mg anti-CD40 per mL of 0.2 mM mannan-BAM in PBS (MBTA therapy).

Uher O., Hadrava Vanova K., Lencova R., Frejlachova A., Wang H., Zhuang Z., Zenka J, & Pacak K. (2023). Intratumoral immunotherapy of murine pheochromocytoma shows no age-dependent differences in its efficacy. Frontiers in Endocrinology, 14, 1030412.

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