Peritoneal macrophages (PM) were collected from Hsf1+/+ and Hsf1−/− mice by peritoneal lavage with Dulbecco’s phosphate-buffered saline as described previously (8 (link)). Thioglycolate-elicited peritoneal cells were obtained from animals 3 days after intraperitoneal injection of 3 mL Thioglycolate broth (Sigma-Aldrich, St. Louis, MO). Macrophages were further purified by adherence to culture dishes for 24 h and identified using anti-HSF1 antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) before use. After 24 h of in vitro culture, peritoneal macrophages were treated with 1,000 ng/mL of LPS for 8 h. RAW264.7 cells (mouse leukemia cells of monocyte macrophage) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured with Dulbecco’s modified Eagle medium (DMEM) plus 10% fetal bovine serum under 5% CO2 conditions at 37°C. RAW264.7 cells were treated with 1,000 ng/mL of LPS for 8 h.
Raw264.7 cells
Manufactured by Shanghai Cell Bank
Sourced in China
RAW264.7 cells are a mouse macrophage cell line derived from the ascites of a tumor induced by the Abelson murine leukemia virus. They are commonly used in cell biology research as a model for macrophage function and inflammation.
Automatically generated - may contain errors
Lab products found in correlation
Thioglycolate, by Merck Group (1 mentions)
Thioglycolate broth, by Merck Group (1 mentions)
Hek293 cells, by Shanghai Cell Bank (1 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
1100 series hplc, by Agilent Technologies (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
2 2 azinobis 3 ethylbenzothiazoline 6 sulfonic acid (abts), by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Meilun (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Shanghai Cell Bank (1 mentions)
Chitosan, by Energy Chemical (1 mentions)
Dnase, by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
Anti neutrophil elastase, by Abcam (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Beyotime (1 mentions)
16 protocols using raw264.7 cells
1
Isolation and Stimulation of Macrophages
2
Culturing U87 Glioma and RAW264.7 Cells
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U87 glioma cells and mouse macrophage RAW264.7 cells were obtained from Cell Bank of Shanghai, Chinese Academy of Sciences (Shanghai, China). The U87 cells were cultured in high-glucose Dulbecco’s modified Eagle medium (DMEM) containing 10% (v/v) FBS and 1% (v/v) antibiotics (penicillin−streptomycin, 100 U/mL). The RPMI 1640 medium was chosen for the maintenance of the RAW264.7 cells. Both abovementioned cells were incubated at 37 °C in 95% air and 5% CO2 incubator.
3
Cell Culture and Animal Care Protocols
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RAW264.7 cells (mice macrophages) and HEK-293 cells (normal human embryonic kidney cells) were bought from the Shanghai Cell Bank of the Chinese Academy of Sciences, Shanghai, China, and cultured with DMEM complete medium. SMMC-7721 cells (human liver cancer cells) and NB4 cells (APL cells) were obtained from Shanghai Jihe Biotechnology Co., Ltd., Shanghai, China and cultured with RPMI 1640 complete medium. Culture of the cells was performed in an incubator kept at 5% CO2 and 37 °C. The male BALB/c nude mice (SCXK 2017-0005) were obtained from Shanghai Slack Laboratory Animals Co., Ltd., Shanghai, China and kept in the Specific Pathogen Free (SPF) animal room of School of Pharmacy, Shanghai Jiao Tong University. Guidelines for care and use of laboratory animals of Shanghai Jiao Tong University were used to perform animal studies and these studies were duly approved by the animal ethics committee of Shanghai Jiao Tong University (No: A2019046, Date: 5 July 2019).
4
Targeted Breast Cancer Therapy Formulation
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DMPC and DOPE were purchased from Shanghai Advanced Vehicle Technology Pharmaceutical Ltd (Shanghai, China). The ApoA1-mimetic peptide (PK-22) was synthesized by GL Biochem (Shanghai) Ltd (Shanghai, China). Mertansine was supplied by BrightGene BioMedical Technology CO. Ltd (Jiangsu, China). DiR was supported by Amyjet Scientifc Inc. (Wuhan, China), and DiI was obtained from Beyotime Institute of Biotechnology (Jiangsu, China). Natural mature HDL from human plasma (lyophilized powder, L1567) was supplied by Sigma-Aldrich (MO, USA).
The murine 4T1 cells, RAW 264.7 cells, and human MCF-7 cells were provided from Cell Bank of Shanghai, Chinese Academy of Sciences (CAS, Shanghai, China). The 4T1-GFP cells were obtained from Keyuandi Biotech Co. Ltd (Shanghai, China). The 4T1, 4T1-GFP, and MCF-7 cells were kept in RPMI-1640 media with 10% FBS (Gibco), and cultured at 37 °C and 5% CO2 in a humidified incubator for further measurements. RAW 264.7 were incubated with Dulbecco’s Modified Eagle Medium (DMEM) with 10% FBS, and cultured as described above. Female nude mice (18–22 g) were provided by Shanghai Experimental Animal Center, CAS (Shanghai, China) and used to induce the breast cancer tumor model. The animal experiments were performed according to the protocols approved by the Institutional Animal Care and Use Committee (IACUC) of Shanghai Institute of Materia Medica, CAS.
The murine 4T1 cells, RAW 264.7 cells, and human MCF-7 cells were provided from Cell Bank of Shanghai, Chinese Academy of Sciences (CAS, Shanghai, China). The 4T1-GFP cells were obtained from Keyuandi Biotech Co. Ltd (Shanghai, China). The 4T1, 4T1-GFP, and MCF-7 cells were kept in RPMI-1640 media with 10% FBS (Gibco), and cultured at 37 °C and 5% CO2 in a humidified incubator for further measurements. RAW 264.7 were incubated with Dulbecco’s Modified Eagle Medium (DMEM) with 10% FBS, and cultured as described above. Female nude mice (18–22 g) were provided by Shanghai Experimental Animal Center, CAS (Shanghai, China) and used to induce the breast cancer tumor model. The animal experiments were performed according to the protocols approved by the Institutional Animal Care and Use Committee (IACUC) of Shanghai Institute of Materia Medica, CAS.
5
Flavonoid Content and Antioxidant Assay
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HPLC-DAD-ESI-MS2 analysis was performed on an Agilent 1100 series HPLC (Palo Alto, CA, USA) and Agilent model Ion Trap mass spectrometer equipped with electrospray ionization ion source (ESI). The content and antioxidant activities of total flavonoids were assayed with T6 ultraviolet and visible spectrophotometer (Beijing Puxi Tongyong Inc., China).
Standard of rutin, dexamethasone (Dex) and gentamicin (HPLC purity>98.0%) were purchased from National Institutes for Food and Drug Control (Shenyang, China). DPPH, ABTS, TPTZ, LPS, dimethyl sulfoxide (DMSO) and 3-(4, 5-dimethylthiazol-2-yl)-2,5 -diphenyltetrazolium bromide (MTT) were bought from Sigma-Aldrich Chemical Company (St. Louis, MO). Other chemicals were of analytical grade.
Standard strains Escherichia coli (ATCC 25922), Salmonella (ATCC 51812), Staphyloccocus aureus (ATCC 25923) and Streptococcus (ATCC 49619) were obtained from American Type Culture Collection.
RAW 264.7 cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China).
Standard of rutin, dexamethasone (Dex) and gentamicin (HPLC purity>98.0%) were purchased from National Institutes for Food and Drug Control (Shenyang, China). DPPH, ABTS, TPTZ, LPS, dimethyl sulfoxide (DMSO) and 3-(4, 5-dimethylthiazol-2-yl)-2,5 -diphenyltetrazolium bromide (MTT) were bought from Sigma-Aldrich Chemical Company (St. Louis, MO). Other chemicals were of analytical grade.
Standard strains Escherichia coli (ATCC 25922), Salmonella (ATCC 51812), Staphyloccocus aureus (ATCC 25923) and Streptococcus (ATCC 49619) were obtained from American Type Culture Collection.
RAW 264.7 cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China).
6
Culturing RAW264.7 Cells for Exosome Isolation
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RAW264.7 cells were purchased from Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, People’s Republic of China). Cells were maintained in DMEM (HyClone) supplemented with 10% FBS (v/v) and 1% penicillin and streptomycin (v/v) at 37°C in a 5% CO2 humidified atmosphere. The conditioned medium for exosome collection was DMEM plus 1% penicillin–streptomycin and 10% FBS pre-centrifuged at 120,000×g for 140 minutes to remove serum exosomes.
7
Cultivation of Murine Macrophage Cell Line
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RAW 264.7 cells belonging to a murine macrophage cell line were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). These cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum, 100 U/ml penicillin, and a streptomycin mixture with supplemented 5% CO2 at 37°C.
8
WB-F344 and RAW264.7 Co-Culture Protocol
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WB-F344 cells (a rat oval cell line that is morphologically and functionally similar to freshly isolated HPCs[23 (link)]) and RAW264.7 cells (a murine macrophage cell line) were purchased from the Shanghai Cell Bank (Chinese Academy of Sciences, Shanghai, China). WB-F344 cells were cultured at 37 °C in an atmosphere containing 5% CO2 in Dulbecco's modified eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS), 2 mmol/L glutamine, and penicillin/streptomycin (100 mg/mL). Activation of RAW264.7 cells was induced with LPS 100 ng/mL for 8 h at 37 °C in an atmosphere containing 5% CO2 in DMEM supplemented with 10% FCS[24 (link)], and then co-cultured with WB-F344 cells in Transwell chambers. A total of 2 × 104 WB-F344 cells were seeded into the upper compartment and 4 × 104 RAW264.7 cells were seeded into the lower compartment of the Transwell chamber. LPS was added to the culture medium, and the medium was replaced every 48 h for a total culture time of 7 d.
9
Culturing RAW264.7 Murine Macrophages
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RAW264.7 cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured in DMEM containing 10% FBS, 100 U/mL penicillin, and 100 U/mL streptomycin. These cells were maintained in a humidified incubator at 37 °C, where CO2 accounted for 5%.
10
Th17 Differentiation and Cell Culture
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CD4+ T cells were isolated from CIA rats and used to differentiate into Th17 cells. The Th17 cells were then cultured in RPMI-1640 medium supplemented with 10% FBS and 1% penicillin-streptomycin. RAW264.7 cells were purchased from Shanghai Cell Bank, Chinese Academy of Sciences (Shanghai, China). HFLS-RA cells were purchased from Huatuo Biotechnology Co. Ltd. (Guangzhou, China). The two cell types were cultured in DMEM (Meilun Biotechnology, Dalian, China) supplemented with 10% FBS and 1% penicillin-streptomycin. All cell cultures were incubated at 37°C and 5% CO2.
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