All chemicals and anhydrous solvents used in this work were purchased from commercial sources and used without further purification. FTIR spectra were recorded in a PerkinElmer Spectrum 400 FT-IR spectrophotometer. 1H and 31P{1H} NMR spectra were recorded on Bruker 400 MHz spectrometer. Elemental analyses were carried out using an Elementar Vario Micro Cube elemental analyzer. ESI-MS analysis was performed using a Bruker Impact ESI-Q-TOF system. Theoretical calculations of PM6 semiempirical molecular orbital method were carried out with Gaussian 09. A549 (human lung cancer cell line), KB (human oral cancer cell line) and HaCaT (human skin keratinocyte cell line) were procured from National Centre for Cell Science (NCCS), Pune. MTT [(3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrasodium bromide] was purchased from SigmaAldrich, USA. Ethidium homodimer-1 in 2 mL pbs, propidium iodide, Ribonuclease A were also purchased from SigmaAldrich.
Hacat
Manufactured by National Centre for Cell Science
Sourced in India
HaCaT is a spontaneously immortalized human keratinocyte cell line derived from normal human skin. It is a well-characterized in vitro model system for the study of keratinocyte biology and skin-related research.
Automatically generated - may contain errors
Lab products found in correlation
Mcf 7, by National Centre for Cell Science (2 mentions)
Vario micro cube elemental analyzer, by Elementar (1 mentions)
Spectrum 400 ftir spectrophotometer, by PerkinElmer (1 mentions)
Ribonuclease a, by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
400 mhz spectrometer, by Bruker (1 mentions)
Hek 293, by National Centre for Cell Science (1 mentions)
Hct 116, by National Centre for Cell Science (1 mentions)
Sodium bicarbonate, by Thermo Fisher Scientific (1 mentions)
Mda mb 231, by National Centre for Cell Science (1 mentions)
Mda mb 468, by National Centre for Cell Science (1 mentions)
Lncap, by National Centre for Cell Science (1 mentions)
Co2 incubator, by Binder (1 mentions)
5 protocols using hacat
1
Synthesis and Characterization of Novel Compounds
2
Culturing Human Cell Lines
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Human embryonic kidney (HEK-293), immortalized human keratinocytes (HaCaT), human breast cancer (MCF-7), and human colorectal (HCT-116) cells were obtained from National Centre for Cell Sciences (NCCS) Pune, India. Cells were grown as monolayer cultures in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS), 1% penicillin–streptomycin (50 μg ml−1), and glutamine (2 mM) in a humidified atmosphere of 5% CO2 at 37 °C in T-25 culture flasks and were sub-cultured twice a week.
3
Culturing Breast Cancer Cell Lines
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Human breast cancer cell lines MDA-MB-231, MDA-MB-468, MCF-7, T47D and keratinocytes HaCaT were purchased from National Centre for Cell Science (NCCS), Pune, India. Human mammary epithelial cells (HMEC) were purchased from Lonza Clonetics, San Diego, USA and cultured in recommended medium. Other cells were grown in Dulbecco’s modification of Eagle’s medium (DMEM) supplemented with 3.7 g Sodium bicarbonate (Invitrogen Corporation, CA), antibiotics (10,000 U/L penicillin and 10 mg/L streptomycin) (Himedia, Mumbai, India) and 10% FBS. Adherent monolayer cell lines were maintained in plastic culture flask and incubated at 37 °C in 5% CO2 in humidified incubator.
4
Culturing of LNCaP and HaCaT cell lines
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In this study, two cell lines were used LNCaP and Ha-CaT and obtained from National Centre for Cell Sciences in Pune, India. LNCaP, a human prostate cancer cell line was cultured at 37°C in a CO 2 incubator in RPMI media with 10% FBS (fetal bovine serum) and 1% antimycotic solution. As a normal control cell line, HaCaT (a human keratinocyte cell line) was used, and it was cultured in DMEM-F12 conditions with 10% FBS (fetal bovine serum) and 1% antimycotic solution.
5
Isolation and Culture of Keratinocytes and Fibroblasts
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Immortalized human epidermal keratinocytes, HaCaT, was obtained from National Centre for Cell Science, Pune, India and normal skin fibroblasts (NSF) were isolated from circumcision samples using regular explant culture techniques45 (link). The cells were cultured in Dulbecco's Modified Eagle Medium supplemented with 10% fetal bovine serum, streptomycin (100 μg/mL), penicillin (100 units/mL), gentamicin (30 μg/mL) and amphotericin B (2.5 μg/mL). The cells were maintained at 37°C in a humidified 5% CO2 incubator (Binder, Germany) in 25 cm2 culture flasks. All the chemicals were procured from Sigma- Aldrich (USA) and are cell culture tested. The tissue collection and the experimental protocols performed, in accordance with the proper guidelines and regulations were approved by the Institutional Ethical Committee of Central Leather Research Institute and prior informed consent for donation of clinical specimens was obtained from all the participants.
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