The NSCLC cell lines, H460 and H2126, were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). The 2 cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% heat-inactivated fetal bovine serum, 100 U/ml penicillin and 100 mg/ml streptomycin (all from HyClone; GE Healthcare, Chicago, IL, USA) at 37°C with 5% CO2. BKM120 (a selective PI3K inhibitor; S2247), LY294002 (a pan-PI3K inhibitor; S1105), MK-2206 (an AKT allosteric inhibitor; S1078), BEZ235 [a dual PI3K/mechanistic target of rapamycin (mTOR) catalytic inhibitor; S1009], PF-2341066 (a MET inhibitor; S1068) and stattic (a STAT3 inhibitor; S7024) were obtained from Selleck Chemicals (Houston, TX, USA), and dissolved in DMSO. Specific small interfering RNAs (siRNAs) for p110α (si-p110α; 100 nM; cat. no. 6359S), AKT (si-AKT; 100 nM; cat. no. 6211S) and MET (si-MET; 100 nM; cat. no. 6618S) were obtained from Cell Signaling Technology Inc. (Danvers, MA, USA). Transfection was performed using Lipofectamine 2000® in Opti-MEM (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA). Cells were transfected with an individual siRNA or treated with an inhibitor for 24 h prior to subsequent experimentation.
H2126
Manufactured by American Type Culture Collection
Sourced in United States
The H2126 is a laboratory instrument designed for culturing and incubating cells. It provides a controlled environment for growing and maintaining cell lines, with precise temperature, humidity, and gas concentration regulation.
Automatically generated - may contain errors
Lab products found in correlation
Bkm120, by Selleck Chemicals (1 mentions)
Bez235, by Selleck Chemicals (1 mentions)
Stattic, by Selleck Chemicals (1 mentions)
Pf 2341066, by Selleck Chemicals (1 mentions)
Ly294002, by Selleck Chemicals (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Mk 2206, by Selleck Chemicals (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
H1299, by American Type Culture Collection (1 mentions)
H1975, by American Type Culture Collection (1 mentions)
H1573, by American Type Culture Collection (1 mentions)
H1437, by American Type Culture Collection (1 mentions)
H1563, by American Type Culture Collection (1 mentions)
3 protocols using h2126
1
NSCLC Cell Lines Modulation by PI3K/AKT Inhibitors
2
Cell Culture and Stimulation Protocol
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16HBE, A549, NHLF, HFLF, H520 and H2126 (Table 1 ) cells were obtained from the American Type Culture Collection (Rockville, MD). 16HBE, NHLF and HFLF were cultured in complete growth media comprised of Dulbecco's modified Eagle's medium (DMEM; Invitrogen) containing 4 mM L-glutamine (Invitrogen), while A549, H520 and H2126 cells were cultured in RPMI 1640 (Invitrogen). All cell-lines were supplemented with 10% heat inactivated fetal bovine serum (GIBCO Invitrogen), 100 IU/ml penicillin and100 µg/ml streptomycin. The cells were maintained in a humidified atmosphere in 5% CO2 at 37°C and were subcultured by incubating with 0.05% trypsin-0.5 mM ethylenediaminetetraacetate (Invitrogen) at a ratio of 1∶3 – 1∶4, weekly.
For cell stimulation purposes, cells were incubated in serum-free media (Invitrogen) as serum has been shown to stimulate B1R expression. Normal cell culture media was replaced with serum-free media for 12 hr prior to the start of stimulation, the cells were then washed once with 1X PBS before being incubated in the absence and presence of the B1R agonist desArg10KD (DAKD)(Sigma Aldrich) at 100 nM and 1 µM or lipopolysaccharide (LPS)(Sigma Aldrich) at 0.1 µg/µl, for 3, 6 and 24 hr.
For cell stimulation purposes, cells were incubated in serum-free media (Invitrogen) as serum has been shown to stimulate B1R expression. Normal cell culture media was replaced with serum-free media for 12 hr prior to the start of stimulation, the cells were then washed once with 1X PBS before being incubated in the absence and presence of the B1R agonist desArg10KD (DAKD)(Sigma Aldrich) at 100 nM and 1 µM or lipopolysaccharide (LPS)(Sigma Aldrich) at 0.1 µg/µl, for 3, 6 and 24 hr.
3
NSCLC Cell Line Collection and Maintenance
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