Cells were diluted to 5.0 x 104 cells per mL, and 20 uL of cell suspension was added to wells of an opaque 384 well plate. After overnight recovery, cells were subjected to a dose response curve of increasing drug concentration in half log steps. For single drug curves, osimertinib (Cayman AZD9291) range was .00003 to 30 uM. For dual drug curves, osimertinib range was .03 to 10 uM, and either KRAS inhibitor (ARS853, Cayman) or FGFR1 inhibitor (PD166866, Cayman) range was .00003 to 30 uM. 10 mM stocks of drugs were made by diluting with DMSO, and half-log drug series were diluted fresh with complete media. Cell viability was read after 48 hours using Promega’s Cell Titer Glo. Luminescence values were normalized to non-drug treated controls, and plotted using Graphpad.
Osimertinib
Manufactured by Cayman Chemical
Sourced in United States
Osimertinib is a laboratory compound supplied by Cayman Chemical. It is a tyrosine kinase inhibitor.
Automatically generated - may contain errors
Lab products found in correlation
Erlotinib, by Cayman Chemical (3 mentions)
Hcc827, by American Type Culture Collection (2 mentions)
Pc9 cell line, by European Collection of Authenticated Cell Cultures (1 mentions)
Rpmi medium, by Fujifilm (1 mentions)
Pcr mycoplasma detection kit, by Takara Bio (1 mentions)
Gefitinib, by Santa Cruz Biotechnology (1 mentions)
Afatinib, by Santa Cruz Biotechnology (1 mentions)
Neratinib, by Santa Cruz Biotechnology (1 mentions)
Dacomitinib, by Cayman Chemical (1 mentions)
Lapatinib, by Santa Cruz Biotechnology (1 mentions)
Ars 853, by Cayman Chemical (1 mentions)
Celltiter glo, by Promega (1 mentions)
Trypan blue, by Corning (1 mentions)
Gefitinib, by Cayman Chemical (1 mentions)
Celltiter glo ctg reagent, by Promega (1 mentions)
96 well plate, by Corning (1 mentions)
Tc20 automated cell counter, by Bio-Rad (1 mentions)
Paclitaxel, by Cayman Chemical (1 mentions)
Afatinib, by Selleck Chemicals (1 mentions)
Gefitinib, by Fujifilm (1 mentions)
8 protocols using osimertinib
1
Combinatorial Drug Screening in Cancer
2
NSCLC Cell Lines and Targeted Drugs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HCC827 and H1975 cell lines (human NSCLC cell lines) were obtained from the American Type Culture Collection (ATCC), and the PC‐9 cell line (a human NSCLC cell line) was obtained from the European Collection of Authenticated Cell Cultures (ECACC). The PC9‐COR cell line was established from the PC‐9 cell line as previously reported.31 All cell lines were authenticated using the short tandem repeat method and were maintained in RPMI medium (FUJIFILM Wako Pure Chemical Corporation) supplemented with 10% fetal bovine serum (FBS; Cytiva, Marlborough, MA). The EGFR status of the cell lines is summarized in Table S2 . All cell lines were used after they were confirmed to be negative for Mycoplasma contamination with a PCR Mycoplasma Detection Kit (TaKaRa Bio) according to the manufacturer's instructions. Erlotinib and osimertinib were obtained from Cayman Chemical Company. TAS‐121 and TAS‐116 were kindly provided by Taiho Pharmaceutical Co., Ltd.
3
Osimertinib Efficacy in NSCLC Xenografts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H2228 and H1975 cell lines were injected into the left flank of 6–12 week old nu/nu mice. After tumors reached 6mm in the largest dimension, mice were randomized into treatment groups; vehicle control, 25 mg/kg Osimertinib, or 50 mg/kg Osimertinib. The 50 mg/kg dose was only used for mice with H2228 tumors. Osimertinib (AZD9291 Cayman) was diluted using the following in order to achieve a final ratio: 5% DMSO, 40% polyethylene glycol, 5% tween-80, 50% H2O. Max volume of treatment was 10 uL for 1 gram of body weight. Mice were weighed on first day of treatment, and volume of drug was adjusted to achieve proper dose.
4
Inhibition of ErbB Kinase Activity in Embryos
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To block ErbB kinase activity in the embryos, 10 µM of tyrosine kinase inhibitors AG 1478 (Calbiochem), erlotinib, gefitinib, lapatinib, afatinib, neratinib, canertinib (all from Santa-Cruz Biotechnologies), osimertinib, and dacomitinib (both from Cayman Chemical), diluted in DMSO, or DMSO alone was applied in E3 medium at 8 hpf, except as indicated for experimentation shown in Figure 4 D. The final concentration of DMSO was 0.1% of the culture medium.
5
Evaluating EGFR Inhibitor Cytotoxicity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested at 70–80% confluence, stained with trypan blue (Corning, USA), and counted with a TC20 Automated Cell Counter (Bio-Rad, USA). Luminescent based cell viability assays using CellTiter-Glo (CTG) reagent (Promega, USA) were performed in 96 well plate (Corning, USA). A total number of 3,000 cells were plated in of complete medium per well in three replicate per drug concentration with Multidrop reagent dispenser (Thermo Fishers, USA). After 3hrs of incubation, of gefitinib, osimertinib and erlotinib (Cayman, USA) diluted in complete RPMI-1640 medium were added to the cells. Compounds were tested in a threefold dilution in a range of and for gefitinib, osimertinib and erlotinib respectively. After 72hrs of incubation, reagent was add to the cells; incubated for 10 minutes at room temperature and luminescence was measured.
6
Tumor Cell Treatment Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
On day 0, 2 × 105 tumor cells were incubated in a T25 flask. Cells were treated either with 20 nM osimertinib (Cayman Chemical), 200 nM gefitinib (FUJIFILM Wako Pure Chemical Corporation), 400 nM afatinib (Selleck Chemicals LLC), or 3.5 nM paclitaxel (Cayman Chemical) for 72 h. Tumor cells and culture supernatants were collected and used for subsequent experiments.
7
Pharmacological Compounds for In Vitro and In Vivo Studies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the in vitro experiments, Erlotinib (10483), Afatinib (11492), Osimertinib (16237), Thalidomide (14610), Prednisolone (20866), Dexamethasone (11015) were obtained from Cayman Chemical. For the in vivo experiments, Prednisone (20677) and Thalidomide (14610) were purchased from Cayman Chemical. Erlotinib (E-4997), Afatinib (A-8644), Lapatinib (L-4899), Vemurafenib (V-2800), Trametinib (T-8123), Imatinib (I-5577), and Infigratinib (I-1500) were purchased from LC Laboratories. For both in vitro and in vivo usages, etanercept (Enbrel) (411175) was purchased from McKesson Medical Supply. Prednisolone is the active metabolite of prednisone. In vivo, prednisone is converted to Prednisolone by the liver. Prednisolone was used for in vitro experiments and prednisone was used for in vivo experiments.
8
Cell Line Cultivation and Drug Treatments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PC‐9 cells were obtained from the European Collection of Authenticated Cell Culture. H1975 and HCC827 cells were obtained from the American Type Culture Collection. These cell lines were maintained in RPMI 1640 medium supplemented with 10% FBS and 100 U/ml penicillin–streptomycin in a humidified atmosphere at 37°C with 5% CO2. Osimertinib was obtained from Cayman Chemical Co. (Ann Arbor). GSI‐XX was obtained from Calbiochem (San Diego).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!