Vero E6 (ATCC CRL-1586) and Vero E6-TMPRSS2 (NIBSC 100978) cells were cultivated in minimal essential medium (MEM) containing 10% fetal bovine serum (PAN Biotech), 100 IU/ml penicillin G and 100 mg/ml streptomycin (Carl Roth). To ensure the selection of TMPRSS2-expressing cells, the medium for Vero E6-TMPRSS2 cells contained an additional 1000 μg/ml geneticin (G418). CaLu-3 cells (ATCC HTB-55) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) with 20% fetal bovine serum (PAN Biotech), 1% non-essential amino acids, 100 IU/ml penicillin G and 100 mg/ml streptomycin (Carl Roth). All cells were cultivated at 37 °C and 5% CO2.
Streptomycin
Manufactured by Carl Roth
Sourced in Germany, United States
Streptomycin is a broad-spectrum antibiotic used in laboratory settings. It is derived from the bacterium Streptomyces griseus and is effective against a variety of gram-positive and gram-negative bacteria. Streptomycin functions by inhibiting protein synthesis in bacterial cells, thereby disrupting their ability to grow and reproduce.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Carl Roth (13 mentions)
Fetal bovine serum (fbs), by PAN Biotech (8 mentions)
Penicillin g, by Carl Roth (7 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Minimum essential medium (mem), by PAN Biotech (4 mentions)
Low glucose dmem, by Thermo Fisher Scientific (3 mentions)
Sb431542, by Bio-Techne (3 mentions)
Vero e6 cell, by American Type Culture Collection (3 mentions)
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Chloramphenicol, by Carl Roth (2 mentions)
Erythromycin, by Carl Roth (2 mentions)
Ampicillin, by Carl Roth (2 mentions)
Medium 199, by Thermo Fisher Scientific (2 mentions)
Turbofect, by Thermo Fisher Scientific (2 mentions)
Dmem medium, by PAN Biotech (1 mentions)
Amphotericin, by Merck Group (1 mentions)
Foetal calf serum, by Thermo Fisher Scientific (1 mentions)
Tigecycline, by Merck Group (1 mentions)
Vancomycin, by Merck Group (1 mentions)
25 protocols using streptomycin
1
Cultivation of SARS-CoV-2 Permissive Cell Lines
2
Cell Culture and Transfection Protocols
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HEK 293T, HeLa, Cos7, and MCF-7 (ATCC, Manassas, USA) were grown in Dulbecco’s modified Eagle’s medium (DMEM, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS, Biochrome, Berlin, Germany), 100 U per mL penicillin and 100 µg per mL streptomycin (both Carl Roth, Karlsruhe, Germany). For HL-1 cells (gift from W.C. Claycomb), Claycomb medium (Sigma-Aldrich), supplemented with 10% FBS (Biochrome), 100 U per mL penicillin, 100 µg per mL streptomycin, 0.1 mM norepinephrine (Carl Roth) and 2 mM l -Glutamine (Carl Roth) was used. All cells were kept at 37 °C and 5% CO2. Transient transfections were performed using TurboFect (Thermo Fischer Scientific) according to the manufacturer’s instructions, and stable cell lines were established using G418 for selection. Inhibitors were used as follows: Z-VAD-FMK (12 h, 100 µM, Promega, Fitchberg, USA), Q-VD-OPh (12 h, 450 nM, Merck Millipore, Billerica, USA), staurosporine (1 to 8 h, 0.5–2 µM, Sigma-Aldrich), caspase-8 inhibitor II (2 h pretreatment, 20 or 50 µM, Calbiochem, Billerica, USA), Doxorubicin (overnight, 0.1 µM–10 µM, Sigma-Aldrich).
3
Equine Dermal Cell Cultivation Protocol
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Equine dermal (ED) cells were grown in DMEM medium (PAN Biotech, Aidenbach, Germany) supplemented with 10% fetal bovine serum (FBS; PAN Biotech) and 100 U ml−1 penicillin (Roth, Karlsruhe, Germany) and 100 μg ml−1 streptomycin (Alfa Aesar, Kandel, Germany) in a 37°C incubator with 5% CO2 atmosphere. Cells were grown to confluency in a 100 mm tissue culture dish, washed with phosphate buffer saline (PBS), trypsinized with 0.25% trypsin supplemented with 2.5 μmol L−1 EDTA and counted in a Neubauer counting chamber.
4
Carp Kidney Cells for Bioassays
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KCB cells grow adherent as a monolayer until confluence and then start to aggregate to dense clusters. Cells were pre-cultivated at 26 ± 1 °C under ambient CO2 conditions in sterile Cellstar® T25 tissue culture flasks (Carl Roth GmbH & Co. KG; CE48.1) using minimal essential medium (Gibco, Thermo Fisher Scientific Inc.; 21,575,097) supplemented with 10% foetal calf serum (Gibco, Thermo Fisher Scientific Inc.; 16,000,044) and 1% non-essential amino acids (Gibco, Thermo Fisher Scientific Inc.; 11,140,035), with 31.25 µg/mL Penicillin (Carl Roth GmbH & Co KG; CN28.1), 50 µg/mL Streptomycin (Carl Roth GmbH & Co. KG; 0236.1) and 50 µg/mL Amphotericin (Sigma Aldrich Corporation; A2942). Due to the poikilothermic nature of the carp, cells can be stored at 4–7 °C for several days up to weeks without recognisable impacts on vitality. This enables shipping of the cells as cell coated plates as components of ready to use test kits. The performance of the cell line according to the conditions above has been validated at GOBIO, Technische Universität Dresden and RWTH Aachen laboratories.
5
Antibiotic Susceptibility Testing Protocol
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Antibiotic MICs were determined by the broth microdilution method according to the CLSI standard guidelines M07-A8, M100-S20, and M45-P (50 , 51 , 53 ). Meropenem, amikacin, gentamicin, linezolid, and tigecycline were purchased from Sigma Aldrich (Germany). Vancomycin and tetracycline were obtained from Merck (Germany). Clindamycin, ciprofloxacin, lincomycin, and penicillin G were obtained from Applichem (Germany). Ampicillin, chloramphenicol, spectinomycin, streptomycin, and erythromycin were purchased from Carl Roth GmbH (Germany). MICs of the quality control strains S. aureus ATCC 29213 and E. coli ATCC 25922 were recorded on each day for all tested antibiotics and fell within the acceptable range prescribed by the CLSI.
6
Colon Cancer Cell Culture Protocol
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Cell culture and maintenance. Human colon carcinoma cell line DLD1 and HT29 cells were obtained from the American Type Culture Collection (Rockville, MD, USA). For the 2D cell culture, cells were cultured in Dulbecco's modified Eagle's medium (DMEM) (HT29 cell line) or RPMI-1640 (DLD1 cell line) cell culture medium (ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (ThermoFisher Scientific), 2 nM glutamine (ThermoFisher Scientific), 100 UI/ml penicillin (ROTH, Karlsruhe, Germany) and 0.1 mg/ml streptomycin (ROTH) at 37˚C in a humidified atmosphere containing 5% CO 2 . For the 3D cell culture, cells were plated in 0.5 mg/ml Ir-ECM (Geltrex™, ThermoFisher Scientific) and cell culture medium upon a layer of agarose in 24-well cell-culture plates as described elsewhere (14) . For all experiments, cells were harvested after 48 h of cultivation.
7
SARS-CoV-2 Variant Propagation in Cell Lines
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Vero E6 (ATCC CRL-1586) and BHK-21 (ATCC CCL-10) cells were grown in minimal essential medium (MEM) containing 10% fetal bovine serum (PAN Biotech), 100 IU/ml penicillin G and 100 μg/ml streptomycin (Carl Roth) at 37°C and 5% CO2. Parental and mutant SARS-CoV-2 were grown in Vero E6 cells. The ancestral SARS-CoV-2 variant B.1 (SARS-CoV-2/München-1.1/2020/929) (Wölfel et al., 2020 (link)) was used as a challenge virus. Virus stocks were stored at −80°C prior to experimental infections.
8
SARS-CoV-2 Animal Challenge Protocol
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The virus isolate BetaCoV/Germany/BavPat1/2020 [15 (link)] T-cell culture passage 3 was used for SARS-CoV-2 animal challenge. The virus was propagated and titrated on Vero E6 cells (ATCC CRL-1586) in minimal essential medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% fetal bovine serum (PAN Biotech, Aidenbach, Germany), 100 IU/mL penicillin G and 100 µg/mL streptomycin (Carl Roth, Karlsruhe, Germany), and stored at −80 °C prior to experimental infections. Genome integrity, specifically the presence of the furin cleavage site, was confirmed by NGS sequencing of virus stocks used for animal experimentation.
9
Detecting and Enumerating Campylobacter
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CCDA plates were supplemented either with nalidixic acid (BioChemica UK Ltd, Billingham, United Kingdom) or streptomycin (Carl Roth GmbH & Co. KG, Karlsruhe, Germany) (1% w/v) to distinguish between the two Campylobacter strains on culture. To detect even low levels of Campylobacter, swabs and samples for qualitative microbiology were initially enriched in Preston broth (Thermo Scientific Inc., Waltham, MA, USA) prior to incubation on CCDA plates as detailed above. The plates were subsequently examined for Campylobacter-like colonies and Campylobacter was confirmed by phase-contrast microscopy or PCR. Results were summarized for all three experiments to show the total percentage of Campylobacter-positive samples per sampling location.
Campylobacter enumeration was performed in duplicates from ten-fold serial dilutions prepared with phosphate buffered saline (PBS). Each dilution step was dispensed onto CCDA plates and incubated as described above. After 48 h, colonies were counted and concentrations calculated according to a standard protocol [7 (link)].
Campylobacter enumeration was performed in duplicates from ten-fold serial dilutions prepared with phosphate buffered saline (PBS). Each dilution step was dispensed onto CCDA plates and incubated as described above. After 48 h, colonies were counted and concentrations calculated according to a standard protocol [7 (link)].
10
Culturing LLC1 Mouse Lung Carcinoma Cells
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The LLC1 mouse Lewis lung carcinoma cell line was obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were cultured at 37°C in a humidified atmosphere containing 5% CO2 with Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Inc.), 2 mM L-glutamine (Thermo Fisher Scientific, Inc.), 100 U/ml penicillin (Carl Roth GmbH Co., KG, Karlsruhe, Germany) and 0.1 mg/ml streptomycin (Carl Roth GmbH Co., KG).
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