In brief, OS cells were seeded into the six‐well plates at a density of 5 × 105 cells/well. After melatonin treatment, the cells were treated with trypsin reagent (Gibco, USA) and the mixture of detached cells was washed using PBS solution twice. Then, the cells were collected and centrifuged at 300 g for 10 min. Next, the pellet was resuspended with 500 μl 0.4% trypan blue solution (Thermo, USA). The cells were maintained in the solution for 3 min and then analysed under an automated cell counter (TC10, BioRad). The blue colour represented the dead cells, and the cell survival rate (%) was shown as the percentage of the live cells compared to total cells.
Trypsin reagent
Manufactured by Thermo Fisher Scientific
Sourced in Ireland
Trypsin is a proteolytic enzyme used in cell culture and biochemical applications. It functions to cleave peptide bonds, facilitating the dissociation of adherent cells from cell culture surfaces or the separation of proteins in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Amikacin, by Thermo Fisher Scientific (2 mentions)
Ciprofloxacin, by Thermo Fisher Scientific (2 mentions)
Calcitonin antibiotic, by Thermo Fisher Scientific (2 mentions)
Amoxicillin, by Thermo Fisher Scientific (2 mentions)
Metronidazole, by Thermo Fisher Scientific (2 mentions)
Gentamicin, by Thermo Fisher Scientific (2 mentions)
Horse serum, by Thermo Fisher Scientific (2 mentions)
Brucella agar, by Thermo Fisher Scientific (2 mentions)
Vitamin k1, by Thermo Fisher Scientific (2 mentions)
Thioglycolate broth, by Thermo Fisher Scientific (2 mentions)
Trypan blue solution, by Thermo Fisher Scientific (1 mentions)
Hek293t cell, by American Type Culture Collection (1 mentions)
Siha cells, by Thermo Fisher Scientific (1 mentions)
Siha cells, by American Type Culture Collection (1 mentions)
Defibrinated sheep blood, by Thermo Fisher Scientific (1 mentions)
Curcumin powder, by Merck Group (1 mentions)
6 protocols using trypsin reagent
1
Melatonin-Induced Cell Survival Assay
2
Antimicrobial Screening of H. perforatum Oil
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H. perforatum oil was produced by the Keroya Company. Brucella agar, vitamin K1, calcitonin antibiotic, defibrinated sheep blood, Thioglycolate broth, horse serum, trypsin reagent, amoxicillin, ciprofloxacin, metronidazole, amikacin, and gentamicin were purchased from Gibco, Ireland.
3
Culturing HEK293T Cells Efficiently
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HEK293T cells were obtained from the American Type Culture Collection (ATCC) (#CRL-3216).
Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (GIBOC, #C11995500CP) supplemented with 10% FBS (GIBCO, #10091148).
Detachment of HEK cells for passaging was performed using the Trypsin reagent (GIBCO, # BS-00-0784).
Cells were cultured at 37°C, 5% CO2 in a humidified atmosphere.
Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (GIBOC, #C11995500CP) supplemented with 10% FBS (GIBCO, #10091148).
Detachment of HEK cells for passaging was performed using the Trypsin reagent (GIBCO, # BS-00-0784).
Cells were cultured at 37°C, 5% CO2 in a humidified atmosphere.
4
SiHa Cell Culture Protocol
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SiHa cells were obtained from the American Type Culture Collection (ATCC) (#HTB-35).
Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (GIBOC, #C11995500CP) supplemented with 10% FBS (GIBCO, #10091148).
Detachment of SiHa cells for passaging was performed using the Trypsin reagent (GIBCO, # BS-00-0784).
Cells were cultured at 37°C, 5% CO2 in a humidified atmosphere.
Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (GIBOC, #C11995500CP) supplemented with 10% FBS (GIBCO, #10091148).
Detachment of SiHa cells for passaging was performed using the Trypsin reagent (GIBCO, # BS-00-0784).
Cells were cultured at 37°C, 5% CO2 in a humidified atmosphere.
5
HeLa Cell Culture Protocol
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HeLa cells were obtained from the American Type Culture Collection (ATCC) (#CRM-CCL-2).
Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (GIBOC, #C11995500CP) supplemented with 10% FBS (GIBCO, #10091148).
Detachment of HeLa cells for passaging was performed using the Trypsin reagent (GIBCO, # BS-00-0784).
Cells were cultured at 37°C, 5% CO2 in a humidified atmosphere.
Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (GIBOC, #C11995500CP) supplemented with 10% FBS (GIBCO, #10091148).
Detachment of HeLa cells for passaging was performed using the Trypsin reagent (GIBCO, # BS-00-0784).
Cells were cultured at 37°C, 5% CO2 in a humidified atmosphere.
6
Antimicrobial Compound Extraction Protocol
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Curcumin powder was obtained from Sigma-Aldrich (Oakville, ON, Canada), and ethanol and hexane were purchased from Merck Company (Darmstadt, Germany). Thioglycolate broth, Brucella agar, defibrinated sheep blood, horse serum, vitamin K1, calcitonin antibiotic, trypsin reagent, amoxicillin, metronidazole, ciprofloxacin, amikacin, and gentamicin were purchased from Gibco, Ireland.
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