Large yellow croakers were treated in seawater containing 1000 IU/mL penicillin (Sangon Biotech, China) and 1.2 mg/mL streptomycin (Sangon Biotech, China) for 24 h. The livers were dissected under sterile conditions and collected in cold sterile PBS supplemented with 100 U/ml penicillin and 100 μg/ml streptomycin. After washing with Dulbecco’s modified Eagle’s medium (DMEM/F12) (Biological Industries, Beit Haemek, Israel), liver tissue was minced into 1 mm3 pieces and digested with 0.25% trypsin-EDTA (Gibco, USA) at room temperature for 10 min. The reaction was terminated by adding DMEM/F12 medium containing 15% fetal bovine serum (FBS), and the cell suspension was purified through a cell strainer with 70 μm mesh. The isolated cells were collected in a 15 ml sterile centrifuge tube and centrifuged at 500 g for 10 min at 4°C. Cell pellets were resuspended in complete medium (DMEM/F12 medium containing 15% FBS, 100 U/ml penicillin and 100 μg/ml streptomycin). Primary hepatocytes were seeded into six-well culture dishes at a density of 2×106 cells/well and incubated at 28°C in 5% CO2.
Streptomycin
Manufactured by Sangon
Sourced in China, United States
Streptomycin is a broad-spectrum antibiotic used in laboratory settings. It is a protein synthesis inhibitor that disrupts the function of bacterial ribosomes, effectively inhibiting bacterial growth. Streptomycin is commonly used in microbiological research and cultivation techniques.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Sangon (131 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (46 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (17 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (12 mentions)
Rpmi 1640, by Thermo Fisher Scientific (8 mentions)
Fetal bovine serum (fbs), by Sangon (8 mentions)
Ampicillin, by Sangon (8 mentions)
Amphotericin b, by Sangon (7 mentions)
Streptomycin, by Thermo Fisher Scientific (7 mentions)
Kanamycin, by Sangon (7 mentions)
Mda mb 231, by American Type Culture Collection (6 mentions)
Mcf 7, by American Type Culture Collection (6 mentions)
Penicillin, by Thermo Fisher Scientific (6 mentions)
L glutamine, by Thermo Fisher Scientific (6 mentions)
Ciprofloxacin, by Sangon (5 mentions)
Vancomycin, by Sangon (5 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (5 mentions)
Penicillin g, by Sangon (4 mentions)
Dmem medium, by Thermo Fisher Scientific (4 mentions)
Chloramphenicol, by Sangon (4 mentions)
151 protocols using streptomycin
1
Isolation and Culture of Primary Hepatocytes from Large Yellow Croaker
2
Cell Culture and Transfection Protocol
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HEK293T, U2OS, SW480, SW620,HCT116, ZR-75-30, and HeLa cells were purchased from the American Type Culture Collection (ATCC). HeLa, U2OS, HEK293T, Sirt3−/− MEFs, SW620, SW480 and HCT116 cells were cultured in DMEM/ high glucose medium (HyClone) supplemented with 10% fetal bovine serum (BI), 100 units ml−1 penicillin and 100 µg ml−1 streptomycin (Sangon Biotech). ZR-75-30 cells were cultured in RPMI-1640 (GIBCO) with supplemented with 10% fetal bovine serum (BI), MEM NEAA (GIBCO), 100 units ml−1 penicillin and 100 µg ml−1 streptomycin (Sangon Biotech). Cells were maintained in an incubator at 37 °C, in a humidified atmosphere containing 5% CO2. Cell transfection was performed using PEI (Polysciences) based on a 3:1 ratio of PEI (μg): total DNA (μg). Cell transfection for siRNA was carried out by Lipofectamine RNAiMAX according to the manufacturer’s protocol. Synthetic siRNA oligo nucleotides were obtained commercially from Shanghai Genepharma Co, Ltd. siATG5: 5ʹ-GGTTTGGACGAATTCCAACTTGTTT-3ʹ; siATG7: 5ʹ-GCTTTGGGATTTGACACATTT-3ʹ.
3
Isolation and Culture of T. musculis
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The cecal contents of HVEM−/− mice were harvested into sterile PBS and filtered three times through a 100-μm cell strainer. The filtrate was centrifuged at 200 × g for 5 min at 4°C. The pellet was washed twice with PBS. T. musculis enriched in the pellet was further purified using a 40%/80% Percoll gradient. For T. musculis culture, the isolated T. musculis was suspended with BHI broth (Oxoid, catalog no. CM1135) supplemented with a cocktail of broad-spectrum antibiotics, including 100 mg/mL streptomycin (Sangon Biotech, catalog no. A100382), 100 U/mL penicillin (Sangon Biotech, catalog no. A613460), 50 mg/mL vancomycin (Sangon Biotech, catalog no. A600983), 10 mg/mL ciprofloxacin (Sangon Biotech, catalog no. A600310), 20 mg/mL gentamicin (Sangon Biotech, catalog no. A506614), and 0.5 mg/mL amphotericin B (Sangon Biotech, catalog no. 171375). After suspension, T. musculis was then incubated in an anaerobic workstation (Don Whitley Scientific) at 37°C for 2 days.
4
Xenograft Mouse Model for Cancer Research
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Female NCG (NOD/ShiLtJGpt-Prkdcem26Cd52Il2rgem26Cd22/Gpt) mice were purchased from Jiangsu Gempharmatech. NCG mice were maintained in accordance with the guidelines for laboratory animals approved by the Animal Research Committee of the University of Science and Technology of China and housed in an ultraclean barrier facility. The K562 (RRID:CVCL_0004) cell line was purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The Ho8910-Luc cell line was constructed and cryopreserved in our laboratory [29 (link)]. All cell lines were maintained in modified RPMI medium (HyClone, SH30809.01) supplemented with 10% FBS (Biological Industries, Beit HaEmek, Israel), 100 U/mL penicillin and 100 µg/mL streptomycin (Sangon Biotech, Shanghai, China) at 37 °C in a 5% CO2 incubator. All the cell lines were identified with STR profiling by Genewiz cpmpany (Suzhou, China).
5
Acute Dissociation and Transfection of DRG Neurons
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DRG neurons were acutely dissociated from Sprague–Dawley (SD) rats as previously described [24 (link)]. DRG neurons with diameters of less than 10 mm were selected to study K+ channels, since small-sized DRG neurons tend to express Kv4.3 [8 (link)]. SD rats (Hunan SJA Laboratory Animal Co., Ltd., China) were used according to the guidelines of the National Institutes of Health for Care and Use of Laboratory Animals. The experiments were approved by the Animal Care and Use Committee of Hunan Normal University.
The human embryonic kidney 293 T cell line (HEK293T) was obtained from Shanghai Institute of Cell Biology (Chinese Academy of Sciences, China) and maintained in DMEM (Invitrogen, USA) supplemented with 10% heat-inactivated fetal calf serum (GibcoTM, USA), penicillin (100 U/ml, Sangon biotech, China), and streptomycin (100 μg/ml, Sangon biotech, China). Wild-type Kv4.3 or all of the mutants were transiently co-transfected into HEK293T cells, with eGFP using the LipfectamineTM2000 Reagent (Invitrogen, USA). The cells that had green fluorescence were selected to electrophysiological assays.
The human embryonic kidney 293 T cell line (HEK293T) was obtained from Shanghai Institute of Cell Biology (Chinese Academy of Sciences, China) and maintained in DMEM (Invitrogen, USA) supplemented with 10% heat-inactivated fetal calf serum (GibcoTM, USA), penicillin (100 U/ml, Sangon biotech, China), and streptomycin (100 μg/ml, Sangon biotech, China). Wild-type Kv4.3 or all of the mutants were transiently co-transfected into HEK293T cells, with eGFP using the LipfectamineTM2000 Reagent (Invitrogen, USA). The cells that had green fluorescence were selected to electrophysiological assays.
6
Accumulation of d3-acetylated Proteins
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HEK293T cells were cultured in DMEM medium supplemented with 10% fetal bovine serum (BI), 100 units/mL penicillin, and 100 mg mL−1 streptomycin (Sangon Biotech). In order to accumulate d3‐acetylated proteins, cells were treated with 4 mm d3‐aspirin dissolved in DMSO for 6 h. Six fully confluent 150‐mm dishes of cells were collected for each experiment. The experiments were repeated from three individual samples.
7
Isolation and Culture of Murine NK Cells
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Murine primary NK cells were isolated from Percoll-separated hepatic mononuclear cells by magnetic sorting using an NK cell isolation kit (Miltenyi,130-115-118, Germany). NK cells were sorted as cells that expressed NK1.1+CD3− using flow cytometry, and the purity was higher than > 95%. After purification, NK cells were cultured in a 96 round bottom wells in complete RPMI 1640 medium supplemented with 10% (v/v) foetal bovine serum (Hakata, HB-FBS-500, Japan), 100 units/ml penicillin(Sangon Biotech, A100339, China), 100 µg/ml streptomycin (Sangon Biotech, A100382, China) and interleukin-2 (IL-2, 10 U/ml, R&D,402-ML, USA).
8
Culturing Breast Cell Lines for Research
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The normal breast epithelial cells MCF-10A and the breast cancer cell lines MDA-MB-468, MDA-MB-231 and MCF-7 were obtained from the American Type Culture Collection. MCF-10A cells were cultured in MEGM (Cobioer Biosciences) with 100 ng/ml cholera toxin. MDA-MB-468, MDA-MB-231 and MCF-7 cells were cultured in DMEM (Sangon Biotech Co., Ltd.) supplemented with 10% fetal bovine serum (Sangon Biotech Co., Ltd.) and 1% antibiotics (penicillin 100 U/ml; streptomycin 100 µg/ml; Sangon Biotech Co., Ltd.). All cell lines were cultured in an incubator at 37°C with 5% CO2.
9
Isolation and Culture of Kidney-Derived Macrophages from Mudskippers
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Kidney-derived MO/MФ from mudskippers were isolated and cultured as reported previously (Guan et al., 2017 (link); Shen et al., 2020 (link)). Briefly, fish kidney leukocyte-enriched fractions were obtained using Ficoll-Hypaque PREMIUM (1.077 g/mL) (GE Healthcare, USA). Non-adherent cells were removed by washing, and adherent cells were incubated in complete medium (RPMI 1640, 5% mudskipper serum, 5% fetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, USA), 100 U/mL penicillin, 100 μg/mL streptomycin) at 24 °C with 5% CO2. The HEK293T cells were maintained in Dulbecco’s modified Eagle’s medium (Biochrom AG, Germany) supplemented with 10% (v/v) FBS, 100 U/mL penicillin (Sangon Biotech, China), and 100 μg/mL streptomycin (Sangon Biotech) at 37 °C with 5% CO2. The HEK293T cells were then seeded into 6-well plates to allow growth until 70%–90% confluence on the day of transfection.
10
Culturing Colorectal Cancer Cell Lines
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CRC cell lines, including DLD-1 and SW480, were obtained from ATCC. DLD-1 and SW480 cells were maintained and grown in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Waltham, MA, United States), 100 μg/ml streptomycin (Sangon, Shanghai, China), and 100 U/ml penicillin G (Sangon) in a 37°C incubator containing 5% CO2.
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