Sulforhodamine B and Compound C were purchased from Sigma Aldrich. IOX2 was purchased from Selleckchem. DMEM high-glucose medium, foetal bovine serum (FBS), penicillin, streptomycin and BCA protein assay kits were purchased from the Beyotime Institute of Biotechnology (Jiangsu, China). The primary antibodies were diluted 1:1000 before use, including AMPK (Cat. # sc-25792, Santa Cruz Biotechnologies), p-AMPK (Cat. # sc-33524, Santa Cruz Biotechnologies), HIF-1α (Cat. #113642 Abcam), P-gp (Cat. # sc-55510, Santa Cruz Biotechnologies), p53 (Cat. # 10442-1-AP, Proteintech), Bax (Cat. # sc-7480, Santa Cruz Biotechnologies), Cytochrome c (Cat. # sc-13561, Santa Cruz Biotechnologies), Caspase 9 (Cat. # 842, Cell signaling), Caspase 3 (Cat. # 836, Cell signaling), PARP (Cat. # 1442, Cell signaling) and GAPDH (Cat. # sc-25778, Santa Cruz Biotechnologies). All the chemical compounds were analytically pure reagents.
Foetal bovine serum (fbs)
Manufactured by Beyotime
Sourced in China
Foetal bovine serum is a widely used cell culture supplement derived from the blood of bovine fetuses. It provides a rich source of proteins, growth factors, and other nutrients essential for the growth and maintenance of various cell types in vitro.
Automatically generated - may contain errors
Lab products found in correlation
P ampk, by Santa Cruz Biotechnology (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Caspase 9, by Cell Signaling Technology (1 mentions)
Dmem high glucose medium, by Beyotime (1 mentions)
Sulforhodamine b, by Merck Group (1 mentions)
Compound c, by Merck Group (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Bcl2 associated x (bax), by Santa Cruz Biotechnology (1 mentions)
Cytochrome c, by Santa Cruz Biotechnology (1 mentions)
Hif 1α, by Abcam (1 mentions)
Penicillin, by Beyotime (1 mentions)
Streptomycin, by Beyotime (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Penicillin streptomycin antibiotics, by Beyotime (1 mentions)
Rmpi 1640 culture medium, by Merck Group (1 mentions)
Skov3, by American Type Culture Collection (1 mentions)
Ovcar3, by National Collection of Authenticated Cell Cultures (1 mentions)
A2780, by National Collection of Authenticated Cell Cultures (1 mentions)
Tryple, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
4 protocols using foetal bovine serum (fbs)
1
Mechanism of Anticancer Drug Action
2
Cell Culture of Common Ovarian Cancer Lines
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SKOV3 was purchased from the American Type Culture Collection (Manassas, VA, USA), and ES-2, OVCAR3, 3AO, A2780 were acquired from the Chinese academy of sciences cell bank (Shanghai, China), while Hey were kindly provided by GeneChem (Shanghai, China). All cell lines were maintained in RMPI 1640 culture medium (Sigma, USA) supplemented with 10% foetal bovine serum (Kangwei, China) and 1% penicillin-streptomycin antibiotics (Beyotime, China) in a humidified (5% CO2, 95% air, 37 °C) incubator.
3
Isolation and Induction of Calcification in Mouse Aortic Smooth Muscle Cells
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Primary mouse aortic smooth muscle cells (MAoSMCs) were isolated from 6-week-old C57BL/6 mice as described previously [24 (link)]. MAoSMCs were grown to confluence in Dulbecco’s modified Eagle medium:F12 medium supplemented with 10% foetal bovine serum and 1% penicillin/streptomycin(GM) (Beyotime Institute of Biotechnology, Shanghai, China) at 37 °C in 5% CO2 infusion and humidified air. Cells from passages 3 to 8 were used in all experiments. The medium was replaced every 2 d. To induce calcification, MAoSMCs were maintained in the presence of calcifying medium (CM) containing 10 mM β-glycerophosphate and 3 mM calcium chloride for 7 d, after which alkaline phosphatase (ALP) activity was assessed and western blotting was performed. The cells were treated for 12–14 d for calcium concentration and alizarin red staining.
4
Mouse Oocyte Purification Protocol
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Oocyte purification was performed as described previously [69 (link)]. In brief, mouse ovaries were isolated and digested with 90 μl of TrypLE (12,604,021, Thermo, New York, US) at 37 °C for 10 min under constant shock. Digestion was terminated by adding 10 μl foetal bovine serum (C0235, Beyotime, Shanghai, China). After centrifugation at 1000 rpm for 1 min, the supernatant was discarded, and the cells were treated with 80 μl hypotonic buffer (30 mM Tris–HCl, 17 mM trisodium citrate dihydrate, 50 mM sucrose, 5 mM EDTA, 0.5 mM DTT, pH 8.2) containing proteinase inhibitor (1:100, P1005, Beyotime, Shanghai, China) for 30 min. Slides were pretreated with 20 μl of fixation buffer (1% paraformaldehyde, pH 9.2 with 50 mM boric acid) containing 0.15% Triton X-100 (T9284, Sigma, Missouri, USA) applied evenly on slides in advance. Then, 20 μl aliquots of the cell suspension were dripped onto the slides, which were incubated at 37 °C for 4 h in a humidified box. The samples were left to dry at room temperature. Then, immunofluorescence staining was performed according to the protocol described above.
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