Atlantic salmon kidney cells (ASK) were acquired from the ATCC (ATCC® CRL-2747) (Devold et al., 2000 (link)). Monolayers of ASK were maintained at 15°C in Leibovitz medium (L-15, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco), penicillin (100 U/ml, Gibco), streptomycin (100 μg/ml, Gibco), gentamycin (50 μg/ml, US Biological) and β-mercaptoethanol (55 mM, Bio-Rad, USA). A viral inoculum stock of ISAV genotype HPR35, with a titer of TCID50 = 107/mL, was kindly provided by ADL diagnostics. The virus was propagated in monolayers of 70% confluent ASK cells as described in the viral infection section. All supernatants of the culture (aliquots of virus) were stored at −80°C.
Gentamycin
Manufactured by US Biological
Sourced in United States
Gentamycin is a broad-spectrum antibiotic commonly used in laboratory settings. It is effective against a variety of gram-negative and gram-positive bacteria. Gentamycin functions by inhibiting bacterial protein synthesis, thereby preventing bacterial growth and replication.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Crl 2747, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
β mercaptoethanol, by Bio-Rad (1 mentions)
L glutamine, by Corning (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Celltiter 96 aqueous one solution cell proliferation assay mts, by Promega (1 mentions)
3 protocols using gentamycin
1
Cultivation and Propagation of ISAV in Atlantic Salmon Kidney Cells
2
Atlantic Salmon Macrophage Cell Culture
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SHK-1 is a macrophage cell line isolated from Atlantic salmon head-kidney (Salmo salar, ECACC Number 97111106, European Collection of Cell Culture, UK). These cells were grown in Leibovitz’s 15 media (L-15, Gibco®, Invitrogen®, Carlsbad, CA, USA) supplemented with 4 mM L -glutamine (Corning cellgro®, Mediatech), Fetal Serum Bovine at 10% (Hyclone®, Thermo Fischer Scientific), gentamycin at 50 µg/mL (US Biological) and 40 μM β-mercaptoethanol (ChemCruz®). Cells were grown at 16 ºC and sub-cultured on reaching a confluence between 80% and 90%.
3
MTS Cell Proliferation Assay Protocol
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Cell medium and antibiotics (RPMI 1640) were from Gibco-BRL (Billings, MT, USA). Fetal bovine serum (FBS) was from Biological Industries (Cromwell, CT, USA). The 3-(4,5-dimethylthiazole-2-yl)-5-(3-carboxymetoxyphenyl)-2-(4-sulfophenyl)-2H internal tetrazolium was from Promega (Madison, WI, USA). The CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) was from Promega (USA). Gentamycin was from US biological (Detrick, MD, USA). Trypsin-EDTA was from Hyclone (Logan, UT, USA). Phosphate-buffered saline (PBS) and penicillin-streptomycin were from Gibco BRL (USA). Trypan blue and sodium dodecylsulfate (SDS) were from Winkler ltda. (Santiago, Chile).
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