M3814 was purchased from Medchemexpress (MCE, HY-101570). Paclitaxel and etoposide phosphate were purchased form Selleck Chemicals (S1150) and Meilunbio (MB1102), respectively. For in vitro studies, M3814 and etoposide was initially dissolved in dimethyl sulfoxide (DMSO; Sigma) as 100 mmol/L stock solution and stored at −20 °C. The stock solution was diluted to reach corresponding concentrations and added to cell cultures. Treatment controls were set as the 0.5% DMSO, the same volume as the other groups. For in vivo experiments, M3814 were orderly added with the following solvent: 10% DMSO, 40% PEG400 (Sigma–Aldrich), 5% Tween-80 (Sigma–Aldrich) and 45% saline. Anti-phospho-histone γ-H2AX (Ser139) was purchased from Abcam (ab11174; Cambridge, UK). Anti-P53 antibody was purchased from HUABIO (ET1601-13; Hangzhou, China). Anti-P21 (2947S), anti-DNA-PKcs (12311S) and anti-p-DNA-PK (Ser2056, 68716S) antibodies were purchased from Cell Signaling Technology (CST, MA, USA). Anti-GAPDH antibody was purchased from Santa Cruz Biotechnology (sc-32233; Santa Cruz, CA, USA). Fluorescent secondary antibodies were obtained from Invitrogen.
M3814
Manufactured by MedChemExpress
M3814 is a laboratory reagent produced by MedChemExpress. It is a small molecule compound used for research purposes. The core function of M3814 is to serve as a tool compound for scientific investigations, though its specific applications and intended uses are not included in this factual description.
Automatically generated - may contain errors
Lab products found in correlation
Paclitaxel, by Selleck Chemicals (1 mentions)
Fluorescent secondary antibody, by Thermo Fisher Scientific (1 mentions)
M3814, by Merck Group (1 mentions)
Anti p21 2947, by Cell Signaling Technology (1 mentions)
Anti gapdh antibody, by Santa Cruz Biotechnology (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Peg400, by Merck Group (1 mentions)
Tween 80, by Merck Group (1 mentions)
Nu7441, by MedChemExpress (1 mentions)
Novobiocin, by MedChemExpress (1 mentions)
Bleomycin, by Merck Group (1 mentions)
Dsirna, by Integrated DNA Technologies (1 mentions)
Rucaparib, by MedChemExpress (1 mentions)
Art558, by MedChemExpress (1 mentions)
Nu7441, by Cayman Chemical (1 mentions)
Bix 01294, by Merck Group (1 mentions)
Gsk126, by Merck Group (1 mentions)
Gsk126, by Selleck Chemicals (1 mentions)
Nu7026, by Merck Group (1 mentions)
Ku57788, by MedChemExpress (1 mentions)
5 protocols using m3814
1
M3814 Pharmacological Evaluation Protocol
2
NHEJ Inhibitor Effects on Large Deletions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NHEJ inhibitors M3814 and NU7441 were purchased from MedChemExpress. These compounds were solubilized in DMSO as 2 mM, and 1 mg per mL, respectively. To investigate the role of NHEJ and MMEJ in large deletion formation, we treated RNP-transfected T cells with DMSO control, 2 μM M3814, or 1 μg per mL NU7441. One day later, the cultures were refreshed with the T Cell Expansion Medium containing no inhibitors.
3
POLQ Inhibitor Screening and Validation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Commercially available small molecules used were: M3814 (MedChemExpress, catalog no. HY-101570), Rucaparib (MedChemExpress, catalog no. HY-10617A), AG-14361 (MedChemExpress, catalog no. HY-12032), L67 (MedChemExpress, catalog no. HY-15586), L189 (MedChemExpress, catalog no. HY-15588), ART558 (MedChemExpress, catalog no. HY-141520), novobiocin (MedChemExpress, catalog no. HY-B0425), B02 (Sigma, catalog no. SML0364) and bleomycin (Sigma, catalog no. B8416). All gRNAs, DNA donors and primers were from Integrated DNA Technologies (Supplementary Data 2 ). SiRNAs were the predesigned smart pool-containing siRNAs 485, 1390, 1397 and 2460 (Horizon Discovery, ON-TARGET plus Human POLQ siRNA - SMART 10721) and the siRNA 765 (Integrated DNA Technologies, DsiRNA hs.Ri.POLQ.13.8).
4
CRISPR-Cas9 Induced Knockout Using Small Molecule Inhibitors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DNA-PKcs inhibitor NU7441 (Cayman; diluted 1:1000 from 1 mM stock in dimethylsulfoxide [DMSO]), M3814 (MedChemExpress; diluted 1:1000 from 1 mM stock in DMSO), GSK126 (Selleckchem; diluted 1:2000 from 1 mM stock in DMSO), BIX01294 (Sigma; diluted 1:1000 from 1 mM stock in H2O), or respective solvent-only controls at equal volumes, was added to the cells at the same time when the cells were supplemented with Shield-1 to induce DD-Cas9 or 24 hours prior to nucleofection for GSK126 and BIX01294. DMSO was also present in the experiments in Figures 3 , 4 , 5 , 6 A, 7 , S2 A–S2F, S3 A, S3B, S4 , S5 , and S7 .
5
Optimized Preparation of Biomolecular Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All molecules used in this study are commercially available: M3814 (MedChemExpress HY-101570), KU57788 (MedChemExpress HY-11006), NU7026 (MedChemExpress HY-15719), LTURM34 (MedChemExpress HY-101667), DMNB (ToChris 2088), and SCR7 Pyrazine (Sigma-Aldrich SML1546). Ten millimolar stocks (and 5 mM for NU7026, owing to solubility limitations) were prepared by resuspension in dimethylsulfoxide (DMSO; Sigma-Aldrich D4540).
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!