24h prior to drug treatments, K562 cells were resuspended in fresh (RPMI) medium at a density of 0.6 × 106 cells/ml. On the day of the experiment, cells were recounted, aliquoted in equal cell numbers to T-25 or T-100 ThermoFisher Tissue Culture Flasks (each flask corresponding to one time point) and treated with Triptolide (Sigma-Aldrich, T3652–1MG) or Trichostatin A (Sigma-Aldrich, T8552–1MG). Final concentrations used in our experiments were: 500 nM Triptolide, and 250nM Trichostatin A. All drug treatments were performed for 0 min, 1h, and respectively 4h.
Tissue culture flasks
Manufactured by Merck Group
Sourced in United Kingdom, United States
Tissue Culture Flasks are laboratory equipment designed for the cultivation and maintenance of cells, tissues, or other biological samples in a controlled environment. They provide a sterile, sealed container to support the growth and study of various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Triptolide, by Merck Group (3 mentions)
Trichostatin a, by Merck Group (2 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
96 well plate, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Trypan blue, by Merck Group (1 mentions)
3 4 5 dimethylthiazole 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Uv visible spectrophotometer, by Shimadzu (1 mentions)
Glacial acetic acid, by Merck Group (1 mentions)
Acridine orange, by Merck Group (1 mentions)
Sulforhodamine b srb, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Ethyl acetate, by Merck Group (1 mentions)
Ethidium bromide, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium, by Merck Group (1 mentions)
Gentamycin, by Merck Group (1 mentions)
Trypsin edta, by HiMedia (1 mentions)
Gallic acid, by Merck Group (1 mentions)
5 protocols using tissue culture flasks
1
Triptolide and Trichostatin A Cytotoxicity
2
Cytotoxicity Evaluation of Cancer Cell Lines
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Human breast carcinoma (MCF-7) and colon cancer (HT-29) cell lines were purchased from Pasteur Institute of Iran (Tehran, I.R. Iran). Roswell park memorial institute (RPMI) medium 1640, fetal bovine serum (FBS), streptomycin and penicillin were provided from Gibco Invitrogen Corporation (UK). Pipettes, tissue culture flasks, 96-well plates, trypan-blue, and 3-[4,5-dimethylthiazole-2-yl]- 2,5-diphenyltetrazolium bromide (MTT) were from Sigma-Aldrich Co. (UK). Dimethyl sulfoxide (DMSO) was from Merck (Darmstadt, Germany). UV/visible spectrophotometer (Shimadzu, 2100; Japan).
3
Triptolide and Trichostatin A Cytotoxicity
4
Cytotoxicity Evaluation of Natural Compounds
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Petroleum ether, diethyl ether, butanol and toluene were purchased from Finar Limited, Ahmedabad, India. Ethanol was procured from Hayman Ltd, Essex, UK. Methanol, ethyl acetate and glacial acetic acid were purchased from Merck Specialties Pvt Ltd, Mumbai, India. Dulbecco’s Modified Eagle’s medium, Fetal Bovine Serum, Gentamycin, Sulforhodamine B (SRB), acridine orange, ethidium bromide, Folin Dennis reagent and gallic acid were obtained from Sigma Aldrich, St Louis, USA. Tris base and trypsin-EDTA were purchased from HiMedia Laboratories Pvt. Ltd, Mumbai, India. Trichloroacetic acid and sodium carbonate was purchased from Nice Chemicals Pvt Ltd, Kochi. Tissue culture flasks, multi-well plates and petri plates were obtained from Sigma Aldrich, St Louis, USA.
5
Triptolide-Induced Chromatin Remodeling in K562 Cells
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Cell culture: K562 cells (ATCC, CCL-243) were cultured at 37 o C, 5% CO2 at a density between 0.3-1 x 10 6 cells/mL in RPMI medium (VWR 45000-396) topped up with 10% Fetal Bovine Serum (Genesee Scientific, cat: #25-514). Cells were split at a consistent interval of 3 days, when the cells reach 10 6 cells/mL.
Cell culture for Triptolide time course: 24h prior to Triptolide treatment, K562 cells were resuspended in fresh (RPMI) medium at a density of 0.6 x 10 6 cells/mL. On the day of the experiment, cells were recounted, aliquoted in equal cell numbers to 6 T-100 ThermoFisher Tissue Culture Flasks (each flask corresponding to one time point) and treated with Triptolide (Sigma-Aldrich, T3652-1MG) to a final concentration of 500 nM Triptolide. The Triptolide treatment was performed for 0 min, 15 min, 30 min, 1h, and respectively 4h.
Cells cross-linking for ChIP: After Triptolide treatment, K562 cells were cross-linked in 1% CH2O freshly prepared in 1x PBS on the day of the experiment to reach the final concentration of 0.1% CH2O in the media. Following a 5 min incubation at room temperature on a rocking platform, the cross-linker was quenched with 1M Glycine to reach a final concentration of 0.135 M Glycine. Lastly, cells were washed twice in 1x PBS, then harvested and snap frozen on dry ice.
Cell culture for Triptolide time course: 24h prior to Triptolide treatment, K562 cells were resuspended in fresh (RPMI) medium at a density of 0.6 x 10 6 cells/mL. On the day of the experiment, cells were recounted, aliquoted in equal cell numbers to 6 T-100 ThermoFisher Tissue Culture Flasks (each flask corresponding to one time point) and treated with Triptolide (Sigma-Aldrich, T3652-1MG) to a final concentration of 500 nM Triptolide. The Triptolide treatment was performed for 0 min, 15 min, 30 min, 1h, and respectively 4h.
Cells cross-linking for ChIP: After Triptolide treatment, K562 cells were cross-linked in 1% CH2O freshly prepared in 1x PBS on the day of the experiment to reach the final concentration of 0.1% CH2O in the media. Following a 5 min incubation at room temperature on a rocking platform, the cross-linker was quenched with 1M Glycine to reach a final concentration of 0.135 M Glycine. Lastly, cells were washed twice in 1x PBS, then harvested and snap frozen on dry ice.
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