Hydroxyurea (HU) or MMC was added to culture medium at the indicated concentrations from stock solutions prepared in PBS to induce DNA replication arrest or DNA damage. To inhibit CHK1 activity, we used the UCN-01 compound (ALEXIS biochemicals) at a 600 nM concentration. To accumulate cells in M-phase, nocodazole (Sigma Aldrich) was used at 0,05 μg/ml. RO3306 (Selleck), a CDK1 inhibitor, was prepared in DMSO and added from a stock solution at a concentration of 20 μM. The B02 compound (Selleck), an inhibitor of RAD51 activity, was used at 27 μM. Clorodeoxyuridine (CldU) (Sigma-Aldrich) was dissolved in sterile DMEM as a 200 mM stock solution and used at 20 μM. SLX4, GEN1 and MUS81 expression was knocked-down by transfection with specific siRNAs directed against the coding sequence of the gene (Qiagen) using Interferin siRNAs delivery reagent (Polyplus).
B02 compound
Manufactured by Selleck Chemicals
The B02 compound is a laboratory reagent used for various analytical and research applications. It serves as a key component in a range of chemical processes and experiments. The primary function of the B02 compound is to facilitate specific reactions and analyses within a controlled laboratory environment. No further details on its intended use or application are provided.
Automatically generated - may contain errors
Lab products found in correlation
Ro 3306, by Selleck Chemicals (2 mentions)
Roscovitine, by Selleck Chemicals (2 mentions)
S1 nuclease, by Thermo Fisher Scientific (2 mentions)
5 ethynyl 2 deoxyuridine (edu), by Merck Group (2 mentions)
Nocodazole, by Merck Group (1 mentions)
Specific sirna, by Qiagen (1 mentions)
Camptothecin, by Merck Group (1 mentions)
Chlorodeoxyuridine cldu, by Merck Group (1 mentions)
Iododeoxyuridine, by Merck Group (1 mentions)
Doxycycline, by Merck Group (1 mentions)
Epigallocatechin egc, by Merck Group (1 mentions)
Nu7441, by Selleck Chemicals (1 mentions)
5 protocols using b02 compound
1
DNA Replication Arrest and Damage Induction
2
DNA Replication Inhibitor Assay
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Camptothecin (Sigma-Aldrich) was dissolved in DMSO and a stock solution (10 mM) was prepared and stored at −20°C. Iododeoxyuridine (IdU) and chlorodeoxyuridine (CldU) (Sigma-Aldrich) were dissolved in sterile DMEM to obtain stock solutions of 2.5 mM and 200 mM, respectively, and stored at −20°C. Mirin (Calbiochem), an inhibitor of MRE11 exonuclease activity, was used at 50 μM; the B02 compound (Selleck), an inhibitor of RAD51 activity, was used at 27 mM. The CDK inhibitor roscovitine (Selleck) was used at 20 μM.
3
DNA Replication Regulators Protocol
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HU (Sigma-Aldrich) was added to culture medium at 2 mM or 0.5 mM from a 200 mM stock solution prepared in Phosphate-buffer saline solution (PBS 1X) to induce DNA replication arrest or slowing. RAD52 inhibitor, Epigallocatechin (EGC - Sigma-Aldrich) was dissolved in DMSO at 100 mM and used at 50 μM. The B02 compound (Selleck), an inhibitor of RAD51 activity, was used at 27 μM. CDC7i (XL413, Selleck) was dissolved in sterile water and used 10 μM. Doxycycline (Sigma-Aldrich) was dissolved in DMSO and used 1μ/ml. MIRIN, the inhibitor of MRE11 exonuclease activity (Calbiochem), was used at 50 μM. Polα inhibitor (ST1926 – Sigma-Aldrich) was dissolved in DMSO and used at the concentrations reported on the experiments. S1 nuclease (Invitrogen cat # 18001016) was diluted 1/100 in S1 buffer and used at 20 U/ml. CldU (Sigma-Aldrich) was dissolved in sterile water as a 200 mM stock solution and used at 50 μM. IdU (Sigma-Aldrich) was dissolved in sterile DMEM as a stock solution 2.5 mM and used at 100 for ssDNA and PLA assays or 250 μM for fibers assay. 5-ethynyl-2’-deoxyuridine (EdU) (Sigma-Aldrich) Was dissolved in DMSO and used at 10 μM for EdU incorporation assay or 100 μM for SIRF assay.
4
DNA Replication and Damage Modulation
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HU was added to culture medium at 2 mM from stock solutions 200 mM prepared in Phosphate-buffer saline solution (PBS) to induce DNA replication arrest or DNA damage. The B02 compound (Selleck), an inhibitor of RAD51 activity, was used at 27 µM. CldU (Sigma-Aldrich) was dissolved in sterile water as a 200 mM stock solution and used at 50 μM. IdU (Sigma-Aldrich) was dissolved in sterile DMEM as a stock solution 2.5 mM and stored at −20 °C. RAD52 inhibitor, EGC (Sigma-Aldrich) was dissolved in DMSO at 100 mM, and stock solution was stored at −80° and was used at 50 µM. Mirin, the inhibitor of MRE11 exonuclease activity (Calbiochem), was used at 50 µM. S1 nuclease (Invitrogen cat # 18001016) was diluted 1/100 in S1 buffer and 10 μl aliquots was stocked at 20° and was used at 20 U/ml.
5
Cell cycle inhibitors and DNA damage
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CPT (ENZO Lifesciences) was dissolved in dimethylsulfoxide, and a stock solution (10 mM) was prepared and stored at −20 °C. CDK1 inhibitor RO-3306 (Selleck) was used at final concentration of 9 μM. Mirin (Calbiochem), an inhibitor of MRE11 exonuclease activity, was used at 50 μM; the B02 compound (Selleck), an inhibitor of RAD51 activity, was used at 27 μM. Roscovitine (Selleck), a pan-CDKs inhibitor, was used at final concentration of 20 μM. NU7441 (Selleck), a DNAPKcs inhibitor, was used at final concentration of 1 μM. The WRN helicase inhibitor (NSC 617145) was used at final concentration of 750 nM (ref. 31 (link)). IdU and CldU (Sigma-Aldrich) were dissolved in sterile DMEM at 2.5 and 200 mM, respectively, and stored at −20 °C.
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