The preparation of the plasmids for human α3(IV), α4(IV), and α5(IV) for nanoluciferase complementation system was previously described.13 (link) In brief, the SmBiT and LgBiT fragments were fused to the C-terminal of α3(IV) and α5(IV) in the pFC36K SmBiT TK-neo Flexi and pFC34K LgBiT TK-Neo Flexi vectors (Promega), respectively. α4(IV) was subcloned into pEB multi-Hygromycin vector (Wako) and fused with 3×FLAG tag. Mutants of α3(IV) and α5(IV) were generated by site-directed mutagenesis as previously described.10 (link),13 (link) The plasmid for PPIF/CypD/CypF (HG14689-CM) was from Sino Biological. For stable cell lines, α3(IV)-SmBiT, α4(IV)-3×FLAG, and α5(IV)-LgBiT wild type or G1244D were subcloned into pLVSIN vector (Takara), respectively, containing hygromycin, puromycin, or blasticidin resistance genes. The reagents CsA, PSC-833, and trimethyl amine N-oxide (TMAO) were from Sigma-Aldrich. Dimethyl sulfoxide, glucose, mannitol, sorbitol, sucrose, and taurine were from Nacalai Tesque. Trehalose was from Tokyo Chemical Industry. Brefeldin A (BFA) was from calbiochem. ALV (HY-12559) was obtained from Medchem Express. FK506/Tacrolimus was from Abcam.
Taurine
Manufactured by Nacalai Tesque
Sourced in Japan
Taurine is an organic compound that is a constituent of bile and is also present in the tissues of many animals. It is an amino sulfonic acid that plays a role in various physiological processes.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Nacalai Tesque (1 mentions)
Sorbitol, by Nacalai Tesque (1 mentions)
Sucrose, by Nacalai Tesque (1 mentions)
Mannitol, by Nacalai Tesque (1 mentions)
Trimethylamine n oxide tmao, by Merck Group (1 mentions)
Trehalose, by Tokyo Chemical Industry (1 mentions)
Psc833, by Merck Group (1 mentions)
Alliin, by Merck Group (1 mentions)
6 well plate, by BD (1 mentions)
2 protocols using taurine
1
Plasmid Preparation for Nanoluciferase Assay
2
Neuro2a Cell Growth Analysis
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Mouse neuroblastoma-derived Neuro2a cells were cultured in Dulbecco's Minimal Essential Medium (MEM) supplemented with 50 U/ml streptomycin, 100 U/ml penicillin G and 10% fetal bovine serum at 37°C in an atmosphere of 5% CO 2 . For cell growth analysis, cells were inoculated at 2 × 10 5 cells/well to 6-well plates (BD Falcon, the size of growing area is approximately 10 cm 2 /well) and cultured for 12 h. Then, 10 µM acrolein (Tokyo Chemical Industry, Tokyo, Japan), and 2-furanmethanthiol (Tokyo Chemical Industry), cysteine methyl ester and ethyl ester (Tokyo Chemical Industry), alliin (Sigma-Aldrich), lysine (Nacalai Tesque) and taurine (Nacalai Tesque) at the indicated concentrations, were added to the medium, and cell growth was followed for 3 days. The viability of cells was monitored by staining with 0.2% trypan blue solution.
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