DLD1 BRCA2 (−/−) cells harboring 20 variants of BRCA2 cDNA and empty vector, untransduced DLD1 BRCA2 (−/ −) cells, and DLD1 parental cells were seeded in 96-well plates at a density of 2.0 × 103 cells/well with 100 µl of medium/well, and each drug was added at various concentrations: olaparib (50 nM–5 µM, Selleckchem), niraparib (10 nM–1 µM, Selleckchem), rucaparib (50 nM–5 µM, LC Laboratories), and CBDCA (50 nM–5 µM, Selleckchem). DMSO (Nacalai Tesque) was added to a final concentration of 0.01% (volume/volume) to wells without drugs. Ten microliters of PrestoBlue cell viability reagent (Thermo Fisher Scientific) was added to each well 144 h after exposure to these drugs, and fluorescence intensity was measured with a 2030 ARVO X3 microplate reader and PerkinElmer 2030 Software v4.0 (PerkinElmer) (excitation; 530 nm, emission; 590 nm)55 (link). Wells without cells were assessed as the negative controls, and survival data were graphically analyzed as a sigmoid curve by GraphPad Prism software v8.02 for Mac (GraphPad Software, Inc.).
Cbdca
Manufactured by Selleck Chemicals
Sourced in United States
CBDCA is a laboratory chemical used as a precursor for the synthesis of other chemical compounds. It is a white crystalline solid with a molecular formula of C6H12N2O2. CBDCA is commonly used in various chemical reactions and processes within a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Niraparib, by Selleck Chemicals (1 mentions)
Prism software v8, by GraphPad (1 mentions)
2030 arvo x3 microplate reader, by PerkinElmer (1 mentions)
2030 software v4, by PerkinElmer (1 mentions)
Prestoblue cell viability reagent, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Nacalai Tesque (1 mentions)
Olaparib, by Selleck Chemicals (1 mentions)
Anti gapdh rabbit polyclonal antibody, by Santa Cruz Biotechnology (1 mentions)
2 7 dichlorodihydrofluorescein diacetate h2dcfda d399, by Thermo Fisher Scientific (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Hoechst 33342 dye, by Merck Group (1 mentions)
2 protocols using cbdca
1
Evaluating BRCA2 Variant Sensitivity to PARP Inhibitors
2
Evaluating Cytotoxicity and Apoptosis in HN-3 Cancer Cells
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The HN-3 cell line was obtained from the Asan Medical Center (Seoul, South Korea) (27 (link)). Hoechst 33342 dye, MTT and propidium iodide (PI) were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). CBDCA was purchased from Selleck Chemicals (Houston, TX, USA). Antibodies against cleaved caspase-3 (rabbit, cat. no. AB3623), cleaved poly(ADP-ribose) polymerase (PARP; rabbit, cat. AB3620) were purchased from Merck KGaA (Darmstadt, Germany); goat anti-epidermal growth factor receptor (EGFR; cat. no. sc-03-G; dilution 1:200) and phosphorylated c-Jun (p-c-Jun; cat. no. sc-99) were from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA); rabbit anti-GAPDH polyclonal antibody (cat. no. ab9485; dilution 1:2,000) was supplied by Abcam (Cambridge, UK). 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA; D399) was purchased from Molecular Probes (Thermo Fisher Scientific Inc., Waltham, MA, USA).
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