Cell viabilities were also detected using a real‐time cell analyser (RTCA) as described above. For IC50 calculation, increasing doses of Olaparib (Selleck, #S1060) in 100 μL culture medium was added into cells seeded in an E‐plate for 24 h. After Olaparib treatment for indicated time, cell indexes at the endpoint were used for curve graphing and IC50 calculation. For combined PARPi and STAT3i treatment, 100 μM Olaparib (Selleck, #S1060) and/or 40 μM C188‐9 (Selleck, #S8605) in 100 μL culture medium was added into 100 μL cultured cells after seeded in an E‐plate for 24 h. After treatment for 5 days, cell indexes were normalized to that at 24 h after cells were seeded into E‐plates and were used to determine cell viability.
Olaparib
Manufactured by Selleck Chemicals
Sourced in United States, Germany, United Kingdom, China, France, Japan
Olaparib is a laboratory chemical product. It is a poly(ADP-ribose) polymerase (PARP) inhibitor. Olaparib functions by inhibiting the activity of PARP enzymes, which are involved in DNA repair processes.
Automatically generated - may contain errors
Lab products found in correlation
Talazoparib, by Selleck Chemicals (37 mentions)
Rucaparib, by Selleck Chemicals (35 mentions)
Veliparib, by Selleck Chemicals (32 mentions)
Niraparib, by Selleck Chemicals (27 mentions)
Cisplatin, by Merck Group (25 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (24 mentions)
Cisplatin, by Selleck Chemicals (21 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (21 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (17 mentions)
Etoposide, by Merck Group (15 mentions)
Etoposide, by Selleck Chemicals (12 mentions)
Hydroxyurea, by Merck Group (12 mentions)
Ve 821, by Selleck Chemicals (12 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (12 mentions)
Gemcitabine, by Selleck Chemicals (11 mentions)
Carboplatin, by Selleck Chemicals (11 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (11 mentions)
Ku55933, by Selleck Chemicals (10 mentions)
Aphidicolin, by Merck Group (9 mentions)
Nu7441, by Selleck Chemicals (9 mentions)
453 protocols using olaparib
1
Olaparib and STAT3i Combination Therapy
2
Evaluating Olaparib Efficacy Modifiers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess the effect of BSO (Selleckchem), NAC (Selleckchem), or cell death inhibitors including ferrostatin-1 (Selleckchem) and Z-VAD-FMK (Selleckchem) on the efficacy of olaparib (Selleckchem), cells with appropriate density per well were incubated in triplicate in 6-well or 12-well plates for 24 h. Cells were then treated with DMSO, BSO, NAC, olaparib, or olaparib in combination with BSO or NAC or indicated cell death inhibitors at appropriate concentrations for 48 h. olaparib was then removed, and the medium was replaced every 48 h with fresh medium containing DMSO, BSO, NAC, or indicated cell death inhibitors. Cells were stained with 0.5% crystal violet (Sigma) dissolved in 20% methanol following 1–2 weeks of incubation, after which the number of colonies were counted visually (or crystal violet was redissolved in methanol and then absorbance was measured at 570 nm). The survival fraction was calculated using GraphPad Prism 6 and normalized to that of control (DMSO) cells.
3
Olaparib and Cisplatin Cytotoxicity Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Olaparib (ASD2281, Selleck Chemicals LCC, Houston, TX) was dissolved in molecular-grade water at a concentration of 10 mM. For cell viability assays, the measurements were performed after treatment with different concentrations (0, 5, 10, 15, 20, 30, 40, and 50 µM) of Olaparib for 48 h, as previously described [12 (link), 18 (link)]. For specific assays, cell lines were incubated with Olaparib 10 µM for 24 and 48 h. Cisplatin (S116650MG, Selleck Chemicals) was dissolved in molecular-grade water at a concentration of 10 mM. For specific assays, cell lines were incubated with Cisplatin for 12 and 24 h.
4
Evaluating Drug Sensitivity in Leukemia Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For drug-sensitivity assays, primary BM pro-B/pre-B cells, WT Em-Myc lymphoma cells or Hoxa9-Meis1 AML cells, or their derivatives, were seeded into 96-well plates at a density of 1 Â 10 5 cells/well and treated with serial diluted drugs (5-point 1:4 dilution), including olaparib, niraparib (Selleckchem, S2741), veliparib (Sellechchem, S1004), S63845, etoposide (Selleckchem, S1225), RG-7388. Cell death was assessed at 48 hours using propidium iodide (PI; Sigma, P4864) uptake, or PI/AnnexinV-APC (WEHI mAb lab), measured on an LSRIIW flow cytometer (BD Biosciences). For CellTiter-Glo assays, 1 Â 10 4 cells were seeded into white 96-well plate (Greiner, 655083) and cell viability was assessed using the CellTiter-Glo Luminescent Assay (Promega, G9241).
To test the interaction between olaparib and BML-277 (Selleckchem, S8632), primary pro-B/pre-B cells (1 Â 10 4 ), Em-Myc lymphoma cells (1 Â 10 4 ), or human cancer cell lines (2 Â 10 3 ) were seeded into 96-well white plates and treated with both drugs, either individually, or in a combination matrix (each drug 5-point 1:4 dilution, olaparib horizontal dilution, BML-277 vertical dilution). Cell viability was determined at 48 hours (pro-B/pre-B or Em-Myc) or at 96 hours (human cancer cell lines) using Cell Titer Glo Luminescent Assay. Bliss scores were calculated as described previously (36) .
To test the interaction between olaparib and BML-277 (Selleckchem, S8632), primary pro-B/pre-B cells (1 Â 10 4 ), Em-Myc lymphoma cells (1 Â 10 4 ), or human cancer cell lines (2 Â 10 3 ) were seeded into 96-well white plates and treated with both drugs, either individually, or in a combination matrix (each drug 5-point 1:4 dilution, olaparib horizontal dilution, BML-277 vertical dilution). Cell viability was determined at 48 hours (pro-B/pre-B or Em-Myc) or at 96 hours (human cancer cell lines) using Cell Titer Glo Luminescent Assay. Bliss scores were calculated as described previously (36) .
5
Clonogenic Survival Assay for DNA Damage Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HR and RAD51 focus assays were previously described (Garcin et al. 2019 (link)). RAD51 antibody used for RAD51 focus formation was used 1:1000 (#PC130, EMD Millipore). For clonogenic survival, U2OS cells were seeded at colony forming density (600 cells) on 60-mm dishes (Garcin et al. 2019 (link)). Cells were seeded in triplicate and treated 24 h after plating with either cisplatin or olaparib. Cells were exposed to 0–2 µM cisplatin for one cell cycle, as determined from the doubling time. Cells were continuously exposed to 0–2 µM olaparib (Selleck Chem AZD2281), replacing the media with fresh olaparib every 3 d. Cells were grown for 12–14 d (beginning with treatment day 1) and fixed in 100% methanol. Plates were stained with crystal violet, scanned on a FluorChem M (proteinsimple), and quantified for area density using the Colony Count Analysis Tool (AlphaView SA software). Area density was normalized relative to untreated plates. U2OS and MCF10A RAD51C derivatives were authenticated and deposited in 2019 at the Leibniz Institute DSMZ, German Collection of Microorganisms and Cell Culture (Garcin et al. 2019 (link)).
6
Developing Olaparib-Resistant 22RV1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
22RV1 cells purchased from ATCC were routinely cultured in RPMI1640 medium (Gibco, Grand Island, NY) supplemented with 10% FBS (Atlanta Biologicals, Flowery Branch, GA). To develop olaparib resistant cell line, cells were maintained in the medium supplemented with 10μM of olaparib (Selleck Chemicals, Houston, TX) for 3 months until the viability reached over 95%.
293T cell line (Lenti-X 293T Cell Line) (34 (link)) was ordered from TaKaRa (Cat. No. 632180) cultured by DMEM medium supplemented with 10% Fetal Bovine Serum (FBS). All the cells were cultured at 37°C in 5% CO2 incubator.
For 3D culture, we used Nunclon™ Sphera™ Dishes (Thermo Scientific, Cat. No. 174945) and added methylcellulose (0.5%; Fisher, Cat No. M-352) in RPMI 1640 growth medium supplemented 10% FBS to prevent excessive aggregation of cells in spheroid culture and to maintain even spheroid size.
293T cell line (Lenti-X 293T Cell Line) (34 (link)) was ordered from TaKaRa (Cat. No. 632180) cultured by DMEM medium supplemented with 10% Fetal Bovine Serum (FBS). All the cells were cultured at 37°C in 5% CO2 incubator.
For 3D culture, we used Nunclon™ Sphera™ Dishes (Thermo Scientific, Cat. No. 174945) and added methylcellulose (0.5%; Fisher, Cat No. M-352) in RPMI 1640 growth medium supplemented 10% FBS to prevent excessive aggregation of cells in spheroid culture and to maintain even spheroid size.
7
Radiation Sensitization by CDK and PARP Inhibition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Irradiation was performed at room temperature using the X-RAD 320 iX X-ray tube (Precision X-Ray Inc.; 320 kV, 10 mA, filter: 0.5 mm Cu + 0.5 mm Al, dose rate: 1.2 Gy/min). For inhibition of CDKs, roscovitine (Calbiochem; Stock 20 mM in DMSO) was added in a final concentration of 10 μM, if not mentioned otherwise. PARP-1 was inhibited by olaparib (Selleckchem; Stock: 10 mM in DMSO) in a final concentration of 1 μM. If not stated otherwise, roscovitine was added 24 h and olaparib 2 h prior to irradiation and removed 24 h later. An equal amount of DMSO was used as solvent control.
8
Cytotoxicity Assay for LMS Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LMS cell lines (SK-LMS-1, SK-UT-1, MES-SA, 1×103; SK-UT-1B, 2×103) were seeded in six-well plates, and treatment with dimethyl sulfoxide (DMSO) or olaparib (1–5 µm ; Selleck) was initiated 24 h after seeding and continued for 10 days, with drug replenishment and medium change every 2 days. Pretreatment with cisplatin (5 µm ; Selleck) was performed for 2 h. Thereafter, cells were washed with phosphate buffered saline (PBS) and incubated with DMSO or olaparib as described above. Following drug treatment, cells were fixed with chilled methanol for 10 min, stained with 0.5% crystal violet in 25% methanol for 15 min, and photographed after overnight drying.
9
Olaparib Mitigates Influenza Mortality
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Two grouping methods were applied in this study. For the part of survival rate analysis, mice were randomly divided into 5 groups, that is, the influenza A virus (IAV) group (H1N1 virus + normal saline), the Ola groups (H1N1 virus + 2.5, 5, 10 mg/kg olaparib), and the positive control group (H1N1 virus + 10 mg/kg Oseltamivir). For the part of biochemical detection, mice were divided into 3 groups, the control group (normal saline), the IAV group (H1N1 virus + normal saline), and the IAV + Ola group (H1N1 virus + 10 mg/kg olaparib).
olaparib (Selleck Chemicals, Houston, TX, USA) at different doses was intraperitoneally injected 2 days before and 5 consecutive days after H1N1 virus challenge. The day of virus challenge was defined as experimental day 1. Oseltamivir (Hoffmann-La Roche, Basel, Switzerland) was administered in a similar manner to olaparib.
olaparib (Selleck Chemicals, Houston, TX, USA) at different doses was intraperitoneally injected 2 days before and 5 consecutive days after H1N1 virus challenge. The day of virus challenge was defined as experimental day 1. Oseltamivir (Hoffmann-La Roche, Basel, Switzerland) was administered in a similar manner to olaparib.
10
Establishment of Olaparib-resistant OVCAR3 Cell Line
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
OVCAR3 (ATCC, #HTB‐161), SKOV3 (ATCC, #HTB‐77) and Hey (Pricella, #CL‐0671) cells were cultured in RPMI‐1640 (Gibco, #A1049101) medium. IGROV1 (Sigma, #SCC203), KGN (Pricella, #CL‐0603) and HEK 293 T (ATCC, #CRL‐3216) cells were cultured in DMEM (Gibco, #A11965‐092) medium. PA1 (Cobioer, #CBP60800) cells were cultured in MEM (Gibco, #11095080) medium. All mediums were supplemented with 10% fetal bovine serum (Gibco, #10099‐141C) and 1% penicillin/streptomycin (Gibco, #15140–122). A stable Olaparib‐resistant OVCAR3 (OVCAR3‐R) cell strain was generated by culturing OVCAR3 cells in the continued presence of 12.5 μM Olaparib (Selleck, #S1060) for more than 12 months. All cells were cultured at 37°C in a humidified atmosphere with 5% CO2.
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!