Melanoma cells were plated at a density of 1 × 105 viable cells per well in a 24-well plate and pretreated with 1 mM valproic acid (VPA) for 168 h with a refreshment of VPA-containing medium every 48 h, followed by VPA removal before further treatment. After 168 h, the cells were cultured with 50 nM talazoparib (BMN-673) (MedChemExpress; Cat#HY-16106), and 2 mM dacarbazine (DTIC) (Sigma Aldrich, Burlington, MA, USA) used alone or in combination. A trypan blue exclusion assay was used to determine the viability of the cells after treatments with VPA, BMN-673, and DTIC. Cells were counted within 3 to 5 min of mixing with 0.4% trypan blue using light microscopy with a Neubauer hemocytometer. The experiments were carried out three times, in triplicate.
Bmn 673
Manufactured by MedChemExpress
Sourced in United States
BMN-673 is a potent and selective inhibitor of PARP1 and PARP2 enzymes. It is a lab equipment product suitable for biochemical and cell-based research applications.
Automatically generated - may contain errors
Lab products found in correlation
D luciferin, by Merck Group (1 mentions)
2 2 azobis, by Merck Group (1 mentions)
5 azacytidine, by MedChemExpress (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Rpmi 1640, by Corning (1 mentions)
Azd2281, by MedChemExpress (1 mentions)
Cisplatin, by Merck Group (1 mentions)
5 iodo 2 deoxyuridine, by Merck Group (1 mentions)
5 chloro 2 deoxyuridine, by Merck Group (1 mentions)
Ab6326, by Abcam (1 mentions)
Superfrost plus slides, by Epredia (1 mentions)
Atm 27156 1 ap, by Proteintech (1 mentions)
Realtime glo mt cell viability assay kit, by Promega (1 mentions)
Pf 00477736, by Merck Group (1 mentions)
Prism 7, by GraphPad (1 mentions)
Mk 8776, by Selleck Chemicals (1 mentions)
Olaparib, by Selleck Chemicals (1 mentions)
7 protocols using bmn 673
1
Combination Therapy Enhances Melanoma Cell Viability
2
Cell Viability Evaluation with Cytotoxic Agents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
4‐Vinylbenzyl chloride, 4,4′‐dithiodibutyric acid, triethylamine, 4‐Cyano‐4‐(phenyl‐carbonothioylthio)pentanoic acid, poly(ethylene glycol)methyl ether methacrylate (average Mn = 950), 2,2‐Azobis (isobutyronitrile) (AIBN), Dulbecco's Modified Eagle's Medium (DMEM), RPMI 1640 Medium, trypsin‐EDTA solution, 3‐(4,5dimethylthiazol‐2‐yl)−2,5‐diphenyl tetrazolium bromide (MTT), D‐Luciferin were purchased from Sigma‐Aldrich (MO, U.S.A), BMN673 and 5‐Azacytidine were purchased from MedChemExpress LLC (NJ, USA). Antibodies for western blot, IHC, immunostaining, and flow cytometry were listed in Table S13 (Supporting Information).
3
Ovarian Cancer Cell Line Cultivation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All ovarian cancer cell lines were gifts from Dr. Ronny Drapkin (Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA), and luciferized74 except A431, which was obtained from ATCC (Manassas, VA). All cell lines were cultured in RPMI1640 (Cellgro, Manassas, VA), except COV362 and COV318, which were cultured in DMEM, JHOS2, and Kuramochi, which were cultured in DMEM/Ham's F‐12 50/50. BMN 673 and AZD2281 were purchased from MedChem Express (Princeton, NJ), dissolved in DMSO, and stored in aliquots at −20°C. Cisplatin was purchased from Sigma‐Aldrich (St. Louis, MO).
4
DNA Replication Dynamics under Stress
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For each LCL sample, ∼100 000 cells were labelled with thymidine analogues, 20 µM 5-chloro-2′-deoxyuridine (CldU, Sigma-Aldrich, C6891) for 30 min followed by 200 µM 5-Iodo-2′-deoxyuridine (IdU, Sigma-Aldrich, I7125) for 30 min and subsequently washed thrice with PBS. Labelled cells were mixed with lysis buffer in 1:1 ratio before mounted on SuperFrost Plus slides (Epredia, J1800AMNZ). Cellular DNA were spread by slanting the slides and fixed in 3:1 methanol/acetic acid for 30 min, followed by denaturation with 2.5 N hydrochloric acid for 40 min and subsequently blocked with 2% BSA in PBS-0.1% Tween20 for 40 min at RT. Slides were washed five times with PBS followed by 2 h incubation with 1:200 anti-BrdU/IdU (Becton Dickinson, 347580, mouse) and 1:200 anti-BrdU/CldU (Abcam, ab6326, rat) antibodies. Subsequently, slides were washed five times with PBS and finally stained with anti-mouse AlexaFluor-488 (Abcam, 150157) and anti-rat AlexaFluor-594 (Abcam, 150116) conjugated secondary antibodies for 1 h at RT.
Labelled DNA fibres were imaged with 63× objective (0.103 um/pixel) using a Zeiss Inverted Fluorescence Live Cell Microscope AO7. The experiments were performed in the presence of replication stress inducing agents, such as HydroxyUrea (HU, Sigma-Aldrich, H8627) and PARP inhibitor (PARPi) (Talazoparib BMN-673, MedChem Express), as controls to assess the differences in RF speed.
Labelled DNA fibres were imaged with 63× objective (0.103 um/pixel) using a Zeiss Inverted Fluorescence Live Cell Microscope AO7. The experiments were performed in the presence of replication stress inducing agents, such as HydroxyUrea (HU, Sigma-Aldrich, H8627) and PARP inhibitor (PARPi) (Talazoparib BMN-673, MedChem Express), as controls to assess the differences in RF speed.
5
Glioblastoma Cell Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Glioblastoma cells were seeded at a density of 7 × 103 viable cells per well in a 12 well-plate. Then, the cells were pretreated with 1 mM valproic acid for a duration of 168 h, with a refreshment of VPA-containing medium every 48 h. After a 168-h pretreatment period, the cells were cultured with 50 nM BMN-673 (MedChemExpress; Cat#HY-16106), 6.25 μM TMZ (Sigma-Aldrich, Burlington, MA, USA), either used alone or in combination. To determine cell viability after treatments with VPA, TMZ, and BMN673, a trypan blue exclusion assay was performed. Cells were counted (within 3 to 5 min of mixing with 0.4% trypan blue) by light microscopy, using a Neubauer hemocytometer. The experiments were carried out three times, in triplicate.
6
Investigating DNA Damage Response Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The antibodies GAPDH (#5174), Caspase3 (#9662), PARP (#9532), CyclinB1 (#4135), CDC2 (#9116), p-CDC2 (#9111), CDC25A (#3652), DAN damage kit (#9947) including p-ATM (Ser1981), p-CHK1 (ser345), p-CHK2 (Thr68), phospho-Histone H2AX (Ser139), and 488 Conjugate secondary antibody (#4412) were purchased from CST (Danvers, MA). CHK1 (10362–1-AP), CHK2 (13954–1-AP) and ATM (27156–1-AP) antibodies were purchase from ProteinTech (Rosemont, USA). Anti-Poly (ADP-Ribose) Polymer antibody (ab14459) and Anti-human CD45-FITC (ab134199) antibody was purchased from Abcam (Cambridge, MA). BMN673 was obtained from MedChemExpress (Monmouth Junction, NJ). NL101 was gifted by Hangzhou Minsheng Institute of Pharmaceutical Research (Hangzhou, China).
7
Synergistic effects of CHK1 and PARP inhibitors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded in 96-well plates at low density and treated (48 or 72 h) with the following drugs: CHK1 inhibitors MK-8776 (SCH 900776, Selleckchem, Houston, TX, USA) or PF-00477736 (Sigma Aldrich) at different doses (0.01, 0.1, 1, 10 μM); PARP inhibitors olaparib (AZD2281, Selleckchem, 1, 10 μM) or talazoparib (BMN-673, MedChem express, Monmouth Junction, NJ, USA, 10 μM). Viability assay was performed using the RealTime-Glo MT Cell Viability Assay kit (#G9711, Promega Corporation, Madison, WI, USA), using a Glomax Multidetection Luminometer, according to manufacturer’s protocol. Luminescence data were normalized on controls and plotted as the percentage of viability over the logarithmic concentration of CHK1 or PARP inhibitor. IC50 values were calculated using the GraphPad Prism 7 software (GraphPad, La Jolla, CA). Dose-response matrices and the synergistic score between compounds were obtained using the SynergyFinder tool. The delta scores were calculated as indicated (https://www.synergyfinder.fimm.fi/ ).
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!