The human gastric adenocarcinoma cells BGC-823 and human myeloid leukemia mononuclear cells THP-1 cells were purchased from China Center for Type Culture Collection (CTCC, Wuhan, China). Cells were cultured in Roswell Park Memorial Institute-1640 medium (RPMI-1640, Gibco BRL, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS, Gibco BRL, Grand Island, NY, USA) in a humidified atmosphere (37 °C, 5% CO2). In addition, phorbol-12-myristate-13-acetate (PMA) was purchased from Solarbio (Peking, China).
Phorbol 12 myristate 13 acetate pma
Manufactured by Solarbio
Sourced in China
Phorbol 12-myristate 13-acetate (PMA) is a synthetic compound used in research applications. It functions as a protein kinase C activator.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Sartorius (2 mentions)
Bgc823, by National Collection of Authenticated Cell Cultures (1 mentions)
Thp 1, by National Collection of Authenticated Cell Cultures (1 mentions)
Ho8910pm, by National Collection of Authenticated Cell Cultures (1 mentions)
Skov3, by National Collection of Authenticated Cell Cultures (1 mentions)
Ovcar3, by National Collection of Authenticated Cell Cultures (1 mentions)
Ho8910, by National Collection of Authenticated Cell Cultures (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Fc block, by BD (1 mentions)
Fixation permeabilization kit, by BD (1 mentions)
Golgistop, by BD (1 mentions)
Anti cd3, by BioLegend (1 mentions)
Anti cd4, by BioLegend (1 mentions)
Anti cd8, by BioLegend (1 mentions)
Anti ifn γ, by BioLegend (1 mentions)
Penicillin streptomycin, by Solarbio (1 mentions)
Matrigel, by BD (1 mentions)
Donkey anti rabbit secondary antibody, by Abcam (1 mentions)
6 protocols using phorbol 12 myristate 13 acetate pma
1
Culturing Human Gastric and Myeloid Cells
2
Epithelial OC Cell Line Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The epithelial OC cell lines HO-8910, HO-8910PM, SKOV-3, SKOV-3ip, and OVCAR-3 and the monocytic cell line THP-1 were obtained from the Chinese Academy of Sciences Cell Bank and had been purchased from China Center for Type Culture Collection (CCTCC) and Cell Resource Center, IBMS, CAMS/PUMC (CRC/PUMC). HO-8910 cells stably overexpressing miR-205 and negative control cells were established by the Xiaoying Wu research group of the pathology laboratory of CSU, and the transfection e ciency was veri ed in a previous study [23] . All cell lines were cultured in DMEM or RPMI 1640 medium (Thermo Scienti c, Waltham, MA, USA) supplemented with 10% foetal bovine serum (FBS) (Gibco, Gaithersburg, MD, USA). THP-1 cells (1×10 6 ) were incubated with 100 ng/ml phorbol 12-myristate 13-acetate (PMA) (Solarbio, Beijing, China) for 24-48 h in vitro to induce their differentiation into macrophages. For the exosome treatment experiments, 1 µg/ml exosomes was added to the culture medium of recipient cells (2 × 10 5 ).
3
Quantifying Antigen-Specific T-Cell Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood from the M6-immunized (three doses) mice or rhesus monkeys was collected at 7, 14 and 28 days post-infection. Red blood cells were removed, and the peripheral blood lymphocytes were washed and suspended in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin. Then, the cells were cultured for 5 h in the presence of phorbol 12-myristate 13-acetate (PMA) (50 ng/ml; Solarbio), ionomycin calcium salt (1 μg/ml; Solarbio) and GolgiStop (0.67 μl/ml; BD Biosciences, New Jersey, USA). The cells were then treated with Fc block (mouse or human; BD Biosciences), followed by anti-CD3 (mouse or human; Biolegend, San Diego, CA, USA), anti-CD4 (mouse or human; Biolegend) and anti-CD8 (mouse; Biolegend, human; BD Pharmingen) and subsequently fixed and permeabilized using a fixation/permeabilization kit (BD Pharmingen) and stained with anti-IFN-γ (mouse; Biolegend, human; BD Pharmingen).
4
Macrophage Polarization and Metabolic Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dulbecco's modified Eagle's medium (DMEM), Roswell Park Memorial Institute (RPMI 1640) medium, fetal bovine serum (FBS), 4′,6‐diamidino‐2‐phenylindole (DAPI) stain solution, and 0.25% trypsin‐EDTA solution was purchased from Sangon Biotech. Penicillin–streptomycin and Phorbol 12‐myristate 13‐acetate (PMA) were purchased from Solarbio Science & Technology Co., Ltd. Ammonium fluoride and ethylene glycol (Sinopharm chemical reagent). The cell counting kit‐8 (CCK‐8) solution and Matrigel (BD Biosciences). Lactic acid assay Kit (Colorimetric method) and Glucose assay Kit (O‐toluidine method) were purchased from Lengton Biotechnology. Anti‐Glut1, anti‐HK2, anti‐CD86, anti‐CD206, donkey anti‐rabbit secondary antibodies, anti‐AMPK‐α, and AMPK inhibitors were purchased from Abcam. A medical‐grade Ti rod was purchased from Baoti.
5
Macrophage Polarization and Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
293T/17 cells [purchased from Procell Life Science&Technology Co., Ltd (China)] were cultured in Dulbecco’s Modified Eagle Medium, high glucose, with Sodium Pyruvate (Biosharp, China) containing 10% fetal bovine serum (FBS, BioInd, Israel) at 37℃ in a 5% CO2 humidified cell culture incubator.
THP-1 cells [purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd (China)] were cultured in Roswell Park Memorial Institute Medium (RPMI)-1640 (Gibco, USA) containing 10% fetal bovine serum (FBS, Gibco, USA), 1% v/v penicillin/streptomycin (Gibco, USA) and 1% v/v GlutaMAX (Gibco, USA). To obtain M0 macrophages, THP-1 cells were treated with 10 mg/mL of Phorbol 12-myristate 13-acetate (PMA, Solarbio, China) for 24 h. To induce M0 macrophage polarization into M1 macrophages, M0 macrophages were further treated with 1 µg/mL LPS (Sigma, USA) and 50 ng/mL IFN-γ (Peprotech, USA) for 24 h (Figure S8).
THP-1 cells [purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd (China)] were cultured in Roswell Park Memorial Institute Medium (RPMI)-1640 (Gibco, USA) containing 10% fetal bovine serum (FBS, Gibco, USA), 1% v/v penicillin/streptomycin (Gibco, USA) and 1% v/v GlutaMAX (Gibco, USA). To obtain M0 macrophages, THP-1 cells were treated with 10 mg/mL of Phorbol 12-myristate 13-acetate (PMA, Solarbio, China) for 24 h. To induce M0 macrophage polarization into M1 macrophages, M0 macrophages were further treated with 1 µg/mL LPS (Sigma, USA) and 50 ng/mL IFN-γ (Peprotech, USA) for 24 h (Figure S8).
6
Investigating Cellular Responses with Ruthenium Agents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NaAsO2 was purchased from Sigma-Aldrich (St. Louis, MO). Ruthenium agents were synthesized according to a previous report.33 (link) 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 4-(2-pyridylazo)resorcinol (PAR) and 2-carboxy-2′-hydroxy-5′-sulfoformazylbenzene sodium salt (Zincon) were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). Nitroblue tetrazolium chloride (NBT) and phorbol 12-myristate 13-acetate (PMA) were purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China). RPMI Medium 1640, trypsin–EDTA and fetal bovine serum (FBS) were obtained from Biological Industries (Kibbutz Beit-Haemek, Israel). Anti-PML and anti-CD11b(FITC) antibodies were obtained from Abcam (Cambridge, US). Anti-GAPDH, anti-PARP-1 and anti-caspase-3 antibodies were purchased from Proteintech (Wuhan, China). The apoptosis detection kit and DNA Content Quantitation Assay (Cell Cycle) were purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China). Ultra-purified water was prepared using a Milli-Q Synthesis System (Millipore, Bedford, MA). All other solvents and reagents were used as received.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!