Betulin, betulinic acid, ursolic acid, and vanillin were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Inotodiol was purchased from ALB Technology Ltd. (Henderson, NV, USA). Diaion HP-20, formic acid, glacial acetic acid (99.7%), and perchloric acid (70%) were purchased from Samchun Chemical Co. (Seoul, Korea). Acetonitrile, n-butanol, and methanol were from JT Baker (Phillipsburg, NJ, USA). Cancer cell lines (HT-29, AGS, MCF-7, and PC3) and the macrophage cell line (RAW 264.7) were obtained from Korea Cell Line Bank (Seoul, Korea). Roswell Park Memorial Institute (RPMI) 1640, Dulbecco’s modified Eagle medium (DMEM), and phosphate buffered saline (PBS) were purchased from GIBCO Invitrogen (Grand Island, NY, USA). Fetal bovine serum (FBS) and penicillin/streptomycin were purchased from WelGENE Inc. (Daegu, Korea) and GE Healthcare Life Sciences (South Logan, UT, USA), respectively. DMSO and MTT were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA).
Raw 264
Manufactured by Korean Cell Line Bank
Sourced in United States, Cameroon
RAW 264.7 is a mouse macrophage cell line. It is a commonly used in vitro model for studying macrophage biology and function.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (30 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (16 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (13 mentions)
Dulbecco modified eagle medium (dmem), by Welgene (8 mentions)
Penicillin, by Thermo Fisher Scientific (8 mentions)
Raw264, by Thermo Fisher Scientific (7 mentions)
Streptomycin, by Thermo Fisher Scientific (7 mentions)
Ht 29, by Korean Cell Line Bank (5 mentions)
Hct116, by Korean Cell Line Bank (5 mentions)
Fetal bovine serum (fbs), by Welgene (5 mentions)
Griess reagent, by Merck Group (4 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (4 mentions)
Raw 264, by Welgene (3 mentions)
Raw 264, by Merck Group (3 mentions)
Caco 2, by Korean Cell Line Bank (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Lipopolysaccharides (lps), by Merck Group (3 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (3 mentions)
High glucose dmem, by Thermo Fisher Scientific (3 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions)
104 protocols using raw 264
1
Anticancer and Anti-inflammatory Potential
2
Cell Culture Protocol: Diverse Cell Lines
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3
Culturing Mouse Microglial and Macrophage Cells
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Mouse brain microglial cells (BV-2) were cultured with RPMI 1640 (Hyclone™, GE Healthcare Life Sciences, Logan, UT, USA) containing 10% fetal bovine serum (GE Healthcare Life Sciences) and 1% penicillin-streptomycin solution (Thermo Fisher Scientific). A mouse macrophage cell line (RAW 264.7) was purchased from the Korean Cell Line Bank (KCIB, Seoul, Korea) and cultured with Dulbecco’s Modified Eagle Medium high glucose cultured media (Hyclone™, GE Healthcare Life Sciences), 10% fetal bovine serum (GE Healthcare Life Sciences), and 1% penicillin-streptomycin solution (Thermo Fisher Scientific). The cultured cells were incubated in a humid atmosphere under 5% CO2 at 37 °C.
4
Cell Culture and Macrophage Differentiation
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Lung cancer cell lines A549 and H460 and murine macrophage cell line RAW264.7 were purchased from the Korean Cell Line Bank (Seoul, Korea). A549, H460, and RAW264.7 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Welgene, Gyeongsan, Korea) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific), 100 U/mL penicillin, and 100 U/mL streptomycin at 37 °C with 5% CO2. A549 and H460 cells were used in passage 10–13, and RAW264.7 cells were used in passage 60. BMDMs were isolated from the femur and tibia bones of 6–8-week-old C57BL/6 mice and differentiated after four days in a medium containing macrophage colony-stimulating factor (25 ng/mL; R&D Systems, Minneapolis, MN, USA) at 37 °C with 5% CO2.
5
Immunomodulatory Polymer-Based Nanoparticles
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Poloxamer 407 (pluronic F127), polyvinylpyrrolidone (PVP), dimethyl sulfoxide (DMSO), dichloromethane (DCM), 5-aminofluorescein and dialysis tube were purchased from Sigma-Aldrich. Imiquimod (R837) was purchased from Macrogen (Seoul, Korea). Dulbecco's modified eagle's medium (DMEM)-high glucose, fetal bovine serum (FBS), antibiotics were obtained from Invitrogen (Carlsbad, CA). Cell lysis buffer was used as received. Vectashield antifade mounting medium with DAPI was purchased from Vector Laboratories (Burlingame, CA). Human colorectal carcinoma cell line (HCT116) and murine macrophage cell line (RAW 264.7) were purchased from Korean Cell Line Bank (Seoul, Korea). ELISA kits were purchased from Cusabio (Wuhan, Hubei, China). Six-week-old male BALB/c mice were purchased and bred in a pathogen-free facility at the Pohang University of Science and Technology (POSTECH). All animal procedures were performed in accordance with the Guidelines for Care and Use of Laboratory Animals of POSTECH and approved by the Institutional Animal Care and Use Committee (IACUC). All chemicals were used without further purification.
6
Elemental Composition and Cell Culture Analysis
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The specimens were coated with 0.7 nm of OsO4 (HPC-1SW, Japan). Each specimen was observed using a scanning electron microscope (Hitachi, SU-70) and underwent EDX microanalysis (EDAX Genesis; Pv 77, EDAX, USA) to analyze the elements of the area of interest. The compositions of the elements were compared.
Murine macrophages from the Cell Bank (RAW264.7; Korean Cell Line Bank No. 40071) were grown on the control and experimental discs. The growth of the cell culture was stopped at 1 h after seeding by fixing the samples. All materials including raw materials were prepared for scanning electron microscopic examination. After immobilization of the samples on the plate, each sample was coated with gold and examined using a scanning electron microscope (H-800, Hitachi, Japan).
Murine macrophages from the Cell Bank (RAW264.7; Korean Cell Line Bank No. 40071) were grown on the control and experimental discs. The growth of the cell culture was stopped at 1 h after seeding by fixing the samples. All materials including raw materials were prepared for scanning electron microscopic examination. After immobilization of the samples on the plate, each sample was coated with gold and examined using a scanning electron microscope (H-800, Hitachi, Japan).
7
Nitric Oxide Determination in Macrophages
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For nitric oxide determination, the mouse macrophage cell line RAW264.7 purchased from the Korea Cell Line Bank (Seoul, Korea) was cultured in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin. Cells (1 × 106 cells/mL) in 6 well plates were stimulated with 100 ng/mL LPS for 24 h in the presence of UWG or CWG. Fifty microliters supernatant was incubated with an equal volume of Griess reagent (Sigma) for 15 min at room temperature. We measured the absorbance at 550 nm with the microplate reader. Sodium nitrite was used as a standard.
8
Culturing Murine Macrophages and Human Colon Cancer Cells
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A murine macrophage cell line, RAW 264.7, was purchased from the Korean Cell Line Bank (Seoul, Republic of Korea). These cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% (v/v) bovine serum (Gibco) and 1% penicillin–streptomycin solution (Gibco), followed by incubation at 37 °C in an incubator with 5% CO2 (Thermo Fisher Scientific, Waltham, MA, USA).
Human colon adenocarcinoma cell line, HT-29, was purchased from the Korean Cell Line Bank and cultured in DMEM supplemented with 10% fetal bovine serum (FBS; Corning Inc., Corning, NY, USA) and 1% (v/v) penicillin–streptomycin solution, followed by incubation at 37 °C in an incubator with 5% CO2. The cells were detached from the culture flask using 0.25% trypsin–ethylenediaminetetraacetic acid (Gibco).
Human colon adenocarcinoma cell line, HT-29, was purchased from the Korean Cell Line Bank and cultured in DMEM supplemented with 10% fetal bovine serum (FBS; Corning Inc., Corning, NY, USA) and 1% (v/v) penicillin–streptomycin solution, followed by incubation at 37 °C in an incubator with 5% CO2. The cells were detached from the culture flask using 0.25% trypsin–ethylenediaminetetraacetic acid (Gibco).
9
Murine Macrophage Cytotoxicity Assay
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The murine macrophage cell line RAW 264.7 was purchased from the Korean Cell Line Bank (KCLB, Seoul, Korea). RAW 264.7 cells were cultured in DMEM supplemented with 10% heat-inactivated FBS, streptomycin (100 µg/mL), and penicillin (100 unit/mL) at 37 °C under 5% CO2 humidified incubator. Exponential phase cells were used throughout the experiments. Then, RAW 264.7 cells (1.5 × 104 cells/mL) plated in 24-well plates were fore 16 h and then treated with LPS (1 µg/mL) plus aliquots of the five marine brown algal methanol extract. The cells were then incubated for an additional 24 h at 37 °C under 5% CO2 humidified incubator. After incubation, the cytotoxic assessment was performed using an MTT assay. The formazan crystals were dissolved in dimethyl sulfoxide (DMSO), and the absorbance was measured using an ELISA plate reader at 540 nm (BioTek Instruments, Inc., Winooski, VT, USA). The optical density of the formazan generated in non-treated control cells was considered to represent 100% viability.
10
Cell Culture of Human Lung Cancer and Murine Macrophage Lines
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Human lung cancer cell lines (A549 and H460) and a murine macrophage cell line (RAW 264.7) were obtained from the Korea Cell Line Bank (Seoul, Korea). Cells were cultured in complete Dulbecco's modified Eagle's medium (DMEM; Gibco BRL, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum and 100 U/ml penicillin/streptomycin (Gibco BRL), and maintained in a humidified chamber at 37°C with 5% CO2.
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