To prepare the treated medium, we first created a stock containing 100µM of drug by dissolving Olaparib (AstraZeneca) in 1mL Dimethyl sulfoxide (DMSO), filtering the solution using a 0.22nm syringe filter, and dissolving it in our regular culture medium. Next, we diluted this maximum tolerated dose (MTD) stock with normal culture medium to obtain batches with 1–50µM Olaparib. We verified that the DMSO did not adversely impact the cells’ growth dynamics (not shown).
Olaparib
Olaparib is a lab equipment product manufactured by AstraZeneca. It is a poly(ADP-ribose) polymerase (PARP) inhibitor. The core function of Olaparib is to inhibit the activity of PARP enzymes, which play a role in DNA repair.
Lab products found in correlation
40 protocols using Olaparib
Olaparib Dose-Dependent Growth Assay
To prepare the treated medium, we first created a stock containing 100µM of drug by dissolving Olaparib (AstraZeneca) in 1mL Dimethyl sulfoxide (DMSO), filtering the solution using a 0.22nm syringe filter, and dissolving it in our regular culture medium. Next, we diluted this maximum tolerated dose (MTD) stock with normal culture medium to obtain batches with 1–50µM Olaparib. We verified that the DMSO did not adversely impact the cells’ growth dynamics (not shown).
Dual Drug Treatment with Cisplatin and Olaparib
Cell Culture and PARP1 Inhibition
Evaluating Olaparib and AZD1775 Combination
Olaparib sensitizes cells to radiation
Experimental strategy combining PARP inhibition and cell irradiation.
In the clonogenic assay (top part), cells at 80% confluence were incubated with PARPi 2 h before irradiation, and after 24 h, cells were seeded in 6-well plates. The plates were left at 37 °C for 8 days, and the clones were stained and counted. In the CellTrace analysis (bottom part), cells at 50% confluence were first stained with CellTrace reagent 24 h before being incubated with PARPi. Two hours after PARPi incubation, the cells were irradiated, and after 24 h, they were diluted to allow cell growth without contact inhibition. The flasks were left at 37 °C for 5 days, and the cells were analyzed by flow cytometry.
Cisplatin and PARP1-inhibitor Combination for Cervical Cancer
PARP1-inhibition (PARP1-i) was induced using 4–5 µM olaparib (Lynparza®, AstraZeneca, Cambridge, UK), dissolved in medium continuously.
Combination Therapy for Cancer
Endometrial Cancer Cell Lines and Olaparib Treatment
Histologically, only the HEC-180 cell line was classified as a serous adenocarcinoma; the other cell lines were classified as endometrioid adenocarcinomas. The culture conditions of the 13 endometrial cancer cell lines were described previously [13 (link)]. HEC-180, HEC-251, and HEC-265 cells were maintained in Eagle’s MEM with 10% FBS. HEC-6 cells stably expressing wild-type PTEN were generated by a retroviral infection, as described previously [13 (link)]. Phoenix cells were transfected with retroviral vectors (pFB-neo) that contained tandem affinity purification (TAP)-tagged wild-type PTEN using Lipofectamine 2000 (Invitrogen, Carlsbad, CA) and the resulting supernatants were used to infect HEC-6 cells. Drug selection was used to purify cell populations after infections by neomycin (500 μg/mL, 7 days).
Olaparib (AZD2281/KU0059436) was provided by AstraZeneca (London, UK). Olaparib was solved in DMSO, and the concentration of DMSO in each assay was 0.1%.
Evaluating DNA Damage Responses in Cancer Cells
GFP-CtIP Variant Colony Formation
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