Amphotericin b

Manufactured by Bio&Sell

Sourced in Germany

Amphotericin B is a polyene antifungal medication used in the treatment of serious fungal infections. It is a naturally-occurring compound produced by the bacterium Streptomyces nodosus. Amphotericin B functions by binding to ergosterol, a component of the fungal cell membrane, leading to the formation of pores and disruption of the membrane's integrity, ultimately causing fungal cell death.

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Lab products found in correlation

Ascorbic acid, by Merck Group (3 mentions) Pen strep fungi mix, by Bio&Sell (3 mentions) T25 culture flask, by Sarstedt (3 mentions) Mem amino acid solution, by Merck Group (2 mentions) Dulbecco s modified eagle s medium dmem ham s f12 medium 1 1, by Bio&Sell (2 mentions) Glutamine, by Harvard Bioscience (2 mentions) Foetal calf serum fcs, by Merck Group (2 mentions) Cos 1186w, by Lonza (2 mentions) Cos 1189, by Lonza (2 mentions) Fos 1077, by Lonza (2 mentions) Fos 1140, by Lonza (2 mentions) Fetal bovine serum (fbs), by Bio&Sell (1 mentions) Penicillin, by Bio&Sell (1 mentions) Streptomycin, by Bio&Sell (1 mentions) Penicillin streptomycin solution, by Merck Group (1 mentions)

6 protocols using amphotericin b

Isolation and Expansion of Rabbit Fibroblasts

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Fibroblasts from the LACL were isolated from ACLs of 11 healthy (8 female and 3 male) New Zealand rabbits (approximate 14 months old) derived from the abattoir. Explanted LACLs were sliced into 2 mm² pieces and placed in a T25 culture flask (Sarstedt AG & Co. KG, Nürnbrecht, Germany) with growth medium (Dulbecco’s Modified Eagle’s Medium (DMEM)/Ham’s F12 medium (Bio&SELL GmbH, Feucht, Germany) supplemented with 10% fetal bovine serum (FBS, Bio&SELL GmbH, Feucht, Germany), 1% penicillin/streptomycin solution (Bio&SELL GmbH, Feucht, Germany), 25 µg/mL ascorbic acid (Sigma-Aldrich, Munich, Germany), 2.5 µg/mL amphotericin B (Bio&SELL GmbH, Feucht, Germany) and MEM amino acid solution (Bio&SELL GmbH, Feucht, Germany) for several weeks. Growth medium changes were performed every second to third day. Emigrating cells were harvested with 0.05% trypsin/0.02% ethylenediaminetetraacetic acid (EDTA, Carl Roth GmbH, Karlsruhe, Germany) solution after 7–10 days and could be further expanded up to passage 5.
As a control, 3 native LACLs (mean length of 0.68 ± 0.19 cm) were used in the CyQuant and DMMB Assays (see the description below) and also for the real time PCR analysis (see the description below).

Gögele C., Konrad J., Hahn J., Breier A., Schröpfer M., Meyer M., Merkel R., Hoffmann B, & Schulze-Tanzil G. (2021). Maintenance of Ligament Homeostasis of Spheroid-Colonized Embroidered and Functionalized Scaffolds after 3D Stretch. International Journal of Molecular Sciences, 22(15), 8204.

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Isolation and Culture of Rabbit Cruciate Ligamentocytes

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For isolation of lapine cruciate ligamentocytes anteroir (ACLs) and posterior cruciate ligaments (PCLs) of 10 healthy male and female New Zealand rabbits (between 4 and 24 months old) obtained from the abattoir were harvested. Slices of 1–2 mm of the ACLs and PCLs (Cls) were prepared and transferred into T25 culture flasks (Sarstedt AG & Co. KG, Nürnbrecht, Germany) with growth medium (Dulbecco’s Modified Eagle’s Medium (DMEM)/Ham’s F12 medium (1:1) (Bio&SELL, Feucht, Germany), containing 10% fetal bovine serum (FBS, Bio&SELL), 1% penicillin/streptomycin solution, 25 μg/mL ascorbic acid (Sigma-Aldrich, Munich, Germany), 2.5 μg/mL amphotericin B (Bio&SELL) and MEM amino acid solution (Sigma-Aldrich)).
Growth medium was changed 2–3 times a week. After 7–10 days, CL-derived ligamentocytes emigrated and were detached using 0.05% trypsin/ 0.02% ethylenediaminetetraacetic acid (EDTA) solution (Bio&SELL). The number of viable cells was determined using the trypan blue exclusion assay with a hemocytometer.

Zahn I., Braun T., Gögele C, & Schulze-Tanzil G. (2021). Minispheroids as a Tool for Ligament Tissue Engineering: Do the Self-Assembly Techniques and Spheroid Dimensions Influence the Cruciate Ligamentocyte Phenotype?. International Journal of Molecular Sciences, 22(20), 11011.

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SARS-CoV-2 Neutralizing Antibody Assay

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A Vero cell-based virus-neutralization test (cVNT) was used for measurement of SARS-CoV-2-neutralizing antibodies (see below). Vero cells (CLS Cell Lines Service GmbH, Eppelheim, Germany, order no. 605372) were incubated at 37°C, 5% CO² and 90% humidity in Dulbecco’s modified Eagle’s medium supplemented with 10 % (v/v) fetal calf serum, 3.7 g/L NaHCO₃, 4.5 g/L glucose, 2mM L-glutamine, and 1% (v/v) of Pen-Strep-Fungi mix containing 10,000 U/ml penicillin, 10 mg/mL streptomycin, and 25 μg/mL amphotericin B (all reagents from Bio&SELL GmbH, Feucht, Germany).

Saggau C., Martini G.R., Rosati E., Meise S., Messner B., Kamps A.K., Bekel N., Gigla J., Rose R., Voß M., Geisen U.M., Reid H.M., Sümbül M., Tran F., Berner D.K., Khodamoradi Y., Vehreschild M.J., Cornely O., Koehler P., Krumbholz A., Fickenscher H., Kreuzer O., Schreiber C., Franke A., Schreiber S., Hoyer B., Scheffold A, & Bacher P. (2022). The pre-exposure SARS-CoV-2-specific T cell repertoire determines the quality of the immune response to vaccination. Immunity, 55(10), 1924-1939.e5.

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Canine and Feline Osteosarcoma Cell Culture

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Commercially available canine osteosarcoma cells D-17 (LGC Standards) or primary tumour-derived neoplastic cells (canine: COS_1186w and COS_1189; feline: FOS_1077 and FOS_1140; Lonza) were used. Cells were grown in Dulbecco’s modified eagle’s medium (DMEM) low glucose with 10% foetal calf serum (FCS, Sigma-Aldrich) and 625 pg Amphotericin B (Bio&Sell), 2 nM glutamine (Biochrom) and 1% Pen/Strep/Fungi Mix (Bio&Sell).

Meyer F.R., Steinborn R., Grausgruber H., Wolfesberger B, & Walter I. (2015). Expression of platelet-derived growth factor BB, erythropoietin and erythropoietin receptor in canine and feline osteosarcoma. Veterinary journal (London, England : 1997), 206(1), 67-74.

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Canine and Feline Osteosarcoma Cell Culture

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Isolation of Lapine Cruciate Ligamentocytes

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Lapine cruciate ligaments used for cell isolation were obtained from the abattoir. Lapine cruciate ligamentocytes were isolated from four healthy female and one male New Zealand Rabbits (mean age of 12 months). Surrounding connective tissue was removed and 1–2-mm sized pieces of the cruciate ligaments were prepared and transferred into a T-25 culture flask (Sarstedt AG & Co. KG, Nürnbrecht, Germany) with 2 mL culture medium (Dulbecco’s Modified Eagle’s Medium (DMEM)/Ham’s F12 medium [1:1] (Bio&SELL), containing 10% fetal bovine serum (FBS, Bio&SELL), 1% penicillin/streptomycin solution, 25 μg/mL ascorbic acid (Sigma-Aldrich, Munich, Germany), 2.5 μg/mL amphotericin B (Bio&SELL) and MEM amino acid solution (Sigma-Aldrich)). Culture medium was changed every 2–3 days. After around 1 week, cruciate ligamentocytes emigrated from the explant. Cells were detached after being 80% confluent using 0.05% trypsin/0.02% ethylenediaminetetraacetic acid (EDTA) solution (Bio&SELL). Cell numbers and viability were calculated with a hemocytometer using the trypan blue exclusion assay.

Zahn I., Stöbener D.D., Weinhart M., Gögele C., Breier A., Hahn J., Schröpfer M., Meyer M, & Schulze-Tanzil G. (2021). Cruciate Ligament Cell Sheets Can Be Rapidly Produced on Thermoresponsive poly(glycidyl ether) Coating and Successfully Used for Colonization of Embroidered Scaffolds. Cells, 10(4), 877.

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