Human HEK-293T cells, the human breast cancer cell lines MDA-MB-231, MDA-MB-468, MDA-MB-436, MCF-7, ZR-75-1, SKBR3 and HS578T, and the human normal mammary epithelial cell line MCF10A were purchased from American Type Culture Collection (ATCC, VA, USA). These cell lines were regularly authenticated by STR analysis and checked for mycoplasma contamination. For the culture of MDA-MB-231, MDA-MB-468, HS578T and HEK-293T cells, high glucose DMEM (Macgene, Beijing, China) supplemented with 10% fetal bovine serum (Gemini, CA, USA), 100 U/ml penicillin (Macgene, Beijing, China) and 100 μg/ml streptomycin (Macgene, Beijing, China) were used. MDA-MB-436, MCF-7, ZR-75-1 and SKBR3 cells were maintained in RPMI-1640 (Macgene, Beijing, China) supplemented with 10% fetal bovine serum (Gemini, CA, USA), 100 U/ml penicillin (Macgene, Beijing, China) and 100 μg/ml streptomycin (Macgene, Beijing, China). MCF-10A cells were cultured as previously described [21 (link)]. All of the cells were incubated at 37 ℃ in a humidified atmosphere with 5% CO2. To inhibit RNA or protein synthesis, cells were treated with actinomycin D (Act D, CST, MA, USA) or cycloheximide (CHX, Selleck, TX, USA), respectively, for the indicated periods of time. MG132 (Selleck, TX, USA) was used to inhibit protein degradation via the proteasome pathway.
Streptomycin
Manufactured by Macgene
Sourced in China, United States
Streptomycin is a laboratory equipment product used for the detection and quantification of streptomycin, an antibiotic compound. It is a critical tool for researchers and scientists working in the fields of microbiology, biochemistry, and pharmaceutical development.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Macgene (25 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Dulbecco modified eagle medium (dmem), by Macgene (7 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Rpmi 1640, by Macgene (4 mentions)
Dmem medium, by Macgene (3 mentions)
Wnt7b, by Abcam (2 mentions)
β tubulin and gapdh antibodies, by Solarbio (2 mentions)
Mmp 1, by Abcam (2 mentions)
Wnt5a, by Abcam (2 mentions)
Hek 293t, by Macgene (2 mentions)
High glucose dmem, by Macgene (2 mentions)
Mg132, by Selleck Chemicals (2 mentions)
Mcf 7, by Macgene (2 mentions)
Dulbecco s modified eagle s medium, by Macgene (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Macgene (2 mentions)
1640 medium, by Thermo Fisher Scientific (2 mentions)
Mda mb 468, by Macgene (1 mentions)
26 protocols using streptomycin
1
Cell Culture Conditions for Cancer and Normal Cell Lines
2
Breast Cancer Cell Culture Protocols
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The human TNBC MDA-MB-231 cells, human breast cancer MCF-7 cells and mouse breast cancer 4T1 cells were obtained from Institute of Basic Medical Science, Chinese Academy of Medical Science (Beijing, China). MDA-MB-231 cells were cultured in Dulbecco's modified Eagle medium (DMEM) medium (Macgene, Beijing, China) supplemented with 10% fetal bovine serum (FBS), 100 U mL−1 penicillin, 100 μg mL−1 streptomycin (Macgene, Beijing, China). MCF-7 cells and 4T1 cells were cultured in Roswell Park Memorial Institute (RPMI 1640) (Macgene, Beijing, China) supplemented with 10% FBS, 100 U mL−1 penicillin, 100 μg mL−1 streptomycin. MDA-MB-231 cells and MCF-7 cells were cultured in humidified atmosphere of 5% CO2 at 37 °C, while 4T1 cells were cultured without 5% CO2. The cells for all experiments were in logarithmic phase of growth.
Female BALB/c nude mice of 18–20 g (6–7 weeks old) were purchased from Vital River Laboratory Animal Center (Beijing, China) and maintained in a specific pathogen free (SPF) environment. All animal experiments were carried out in accordance with guidelines, and approved by the Committee for Experimental Animal Welfare of Biomedical Ethics of Peking University.
Female BALB/c nude mice of 18–20 g (6–7 weeks old) were purchased from Vital River Laboratory Animal Center (Beijing, China) and maintained in a specific pathogen free (SPF) environment. All animal experiments were carried out in accordance with guidelines, and approved by the Committee for Experimental Animal Welfare of Biomedical Ethics of Peking University.
3
Cell Line Cultivation and Authentication
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NCI-N87 cells were purchased from American Type Culture Collection (Manassas, VA, USA), SNU-719 cells were purchased from Cobioer (Nanjing, China). HGC-27 cells were purchased from Cell Bank of Chinese Academy of Sciences (Shanghai, China). GES-1 cells were gotten from Peking University Cancer Hospital (Beijing, China). The mycoplasma contamination of all the cell lines was tested as negative. Cell lines were authenticated by short tandem repeat technology. NCI-N87 and SNU-719 cells were cultured in 1640 medium (Gibco-BRL, Grand Island, NY, USA) with 10% fetal bovine serum (Gibco-BRL, Grand Island, NY, USA), 100 U/ml penicillin and 100 μg/ml streptomycin (Macgene, Beijing, China). HGC-27 and GES-1 cells were cultured in DMEM medium (Gibco-BRL, Grand Island, NY, USA) with 10% fetal bovine serum (Gibco-BRL, Grand Island, NY, USA), 100 U/ml penicillin and 100 μg/ml streptomycin (Macgene, Beijing, China). Cells were maintained at 37 °C/5% CO2.
4
Characterization of Cell Lines for Oncology Research
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The human embryonic kidney 293 T (HEK 293 T) and human OSCC cell lines, including FaDu and SCC9, were obtained from the National Platform of Experimental Cell Resources for SCI-Tech (Beijing, China). All cell lines were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, NY, USA), supplemented with 10% fetal bovine serum (FBS; Gibco), 100 U/ml penicillin and 100 μg/ml streptomycin (Macgene, China). Cells were cultured at 37 °C in a humidified incubator under 5% CO2. Primary antibodies used for immunohistochemistry detection and Western blot were as follows: WNT7B (Abcam), WNT5A (Abcam), FZD7 (Abcam) and MMP1 (Abcam), GPC1 (Sigma-Aldrich). β-Tubulin and GAPDH antibodies used for Western blot were obtained from Solarbio Science & Technology Co., Ltd.
5
Silencing of NKp30 in BGC823 and AGS Cells
6
Culturing Mouse BV-2 Microglial Cells
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Mouse BV-2 microglial cells (Peking Union Medical College Cell Bank, Beijing, China) were cultured in Dulbecco’s Modified Eagle’s Medium (Macgene, Beijing, China) and supplemented with 10% fetal bovine serum (Gibico, Waltham, MA, USA), penicillin (Macgene, 100 U/mL, Beijing, China), and streptomycin (Macgene, 100 μg/mL) in a humidified incubator containing 95% air and 5% CO2 at 37 °C.
7
Cell Culture and Recombinant Virus Preparation
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HEK293T (Cat# CRL-11268, ATCC), Vero E6 (Cat# CRL-1586, ATCC), and HeLa (Cat# CCL-2, ATCC) cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco), 2 mmol/L l -glutamine (Gibco), 50 U/mL penicillin, and 50 ug/mL streptomycin (Macgene) at 37 °C in a humidified atmosphere of 5% CO2.
Plasmids encoding EBOV-GP, vesicular stomatitis virus envelope protein (VSV-G) and human immunodeficiency virus type 1 (HIV-l) luciferase reporter vector pNL4-3.Luc.R-E- were kept in the Institute of Medicinal Biotechnology, Beijing, China. The GP genes of MARV (GenBank Accession No.ABA87127.1 , amino acid [a.a.] 5941–7986) were synthesized by Suzhou GENEWIZ Biotechnology Co., Ltd. (Suzhou, China), and inserted into pcDNA3.1(+) vector (Invitrogen, Shanghai, China). Plasmid encoding β-lactamase-Vpr (BlaM-Vpr) was kindly provided by Dr. Xu Tan (Tsinghua University, Beijing, China).
All work with recombinant EBOV expressing enhanced green fluorescent protein (EGFP) (EBOV H.Sapiens-rec/GIN/2014/Makona-Guekedou-C07-EGFP), mouse-adapted EBOV variant Mayinga38 (link) and wild type MARV variant Angola (MARV H.spaiens-tc/AGO/2005/Angola) was performed in the containment level 4 laboratories at the Canadian Science Centre for Human and Animal Health (CSCHAH), Public Health Agency of Canada, Winnipeg, MB, Canada.
Plasmids encoding EBOV-GP, vesicular stomatitis virus envelope protein (VSV-G) and human immunodeficiency virus type 1 (HIV-l) luciferase reporter vector pNL4-3.Luc.R-E- were kept in the Institute of Medicinal Biotechnology, Beijing, China. The GP genes of MARV (GenBank Accession No.
All work with recombinant EBOV expressing enhanced green fluorescent protein (EGFP) (EBOV H.Sapiens-rec/GIN/2014/Makona-Guekedou-C07-EGFP), mouse-adapted EBOV variant Mayinga38 (link) and wild type MARV variant Angola (MARV H.spaiens-tc/AGO/2005/Angola) was performed in the containment level 4 laboratories at the Canadian Science Centre for Human and Animal Health (CSCHAH), Public Health Agency of Canada, Winnipeg, MB, Canada.
8
Macrophage and Chondrogenic Cell Culturing
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RAW264.7 murine macrophages were obtained from Institute of Basic Medical Science, Chinese Academy of Medical Sciences (Beijing, China). The cells were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (PAN, Germany), 100 U mL−1 penicillin, and 0.1 mg mL−1 streptomycin (Macgene, Beijing, China). Cells were cultured in a humidified incubator with 5% CO2 at 37 °C. The cells for all experiments were in the logarithmic phase of growth.
Notably, in this study the “normal macrophages” refers to the RAW264.7 macrophage cells without LPS treatment (M0 macrophages), and the “activated macrophages” refers to the RAW264.7 macrophage cells treated with LPS (5 µg mL−1) for 24 h (M1 macrophages).
Chondrogenic cell ATDC5 was cultured with DMEM (Gibco, USA), containing 100 U mL−1 penicillin, 100 mg mL−1 streptomycin sulfate, and 10% FBS in a humidified incubator with 5% CO2 at 37 °C.
Notably, in this study the “normal macrophages” refers to the RAW264.7 macrophage cells without LPS treatment (M0 macrophages), and the “activated macrophages” refers to the RAW264.7 macrophage cells treated with LPS (5 µg mL−1) for 24 h (M1 macrophages).
Chondrogenic cell ATDC5 was cultured with DMEM (Gibco, USA), containing 100 U mL−1 penicillin, 100 mg mL−1 streptomycin sulfate, and 10% FBS in a humidified incubator with 5% CO2 at 37 °C.
9
RAW264.7 Cell Exposure to TDCPP
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RAW264.7 cells were obtained from the Stem Cell Bank, Chinese Academy of Sciences, and were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin (Macgene, Beijing, China), and 10% fetal bovine serum (Gibco, Waltham, MA, USA) at 37°C in a humid-ified atmosphere containing 5% CO 2 . Cells were treated with various concentrations of TDCPP for 24 hr. Control cells were incubated with 0.1% DMSO to match the final concentration achieved in culture medium in the experimental exposures.
10
Culturing Toxoplasma gondii Tachyzoites
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Cells were cultured in 25 cm2 culture flasks in DMEM medium (Macgene, China) supplemented with 100 U mL−1 penicillin, 100 μg mL−1 streptomycin (Macgene, China) and 10% heat-inactivated foetal bovine serum (FBS) (BI, Israel) at 37°C under a 5% CO2 atmosphere. Toxoplasma gondii tachyzoites (RH strain) were maintained in Vero cells cultured in DMEM medium supplemented with penicillin, streptomycin and 2% FBS at 37°C and 5% CO2.
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