Alanine racemase (Alr) is a bacterial enzyme that catalyses the conversion of L-alanine to D-alanine (D-Ala)1. This function is critical for the growth of bacteria due to their need for D-alanine, an essential component in the biosynthesis of cell wall peptidoglycan in both gram-positive ...
medium: negative control, low control: LDH activity released from the untreated cells (spontaneous LDH release); high control-1: cells were lysed with 5 μl of lysis buffer; high control-2: cells were lysed with 10 μl of lysis buffer; tolytoxin and methanol treatments from 3 nM ...
Oxidation Resistance 1 (OXR1) gene is a highly conserved gene of the TLDc domain-containing family. OXR1 is involved in fundamental biological and cellular processes, including DNA damage response, antioxidant pathways, cell cycle, neuronal protection, a
Vertebrate early embryogenesis is initially directed by a set of maternal RNAs and proteins, yet the mechanisms controlling this program remain largely unknown. Recent transcriptome-wide studies on RNA structure have revealed its pervasive and crucial ro
For luciferase assays, cells were lysed in 60 μl of 1x passive lysis buffer of the Dual-Luciferase Reporter Assay System (Promega, E1980) and directly assayed or frozen at −20°C. After thawing, cell debris and nuclei were removed by centrifugation for 1 min at 13,000 rpm. 20 μl...
Then the cells were incubated on ice before lysed in SDS lysis buffer. The genomic DNA was sonicated in an ultrasonic breaker machine to obtain 200-1000 bp DNA fragments. The cell lysates were added NaCl at 65 °C for four hours to remove cross-links between protein and genomic DNA. The...
Protein extracts from cultured rat hepatocytes were prepared by lysing pre-washed cells with lysis buffer [20 mM HEPES (pH 7.6), 150 mM NaCl, 1 mM EDTA, 2 mM dithiothreitol (DTT) and 1% (v/v) Triton X-100] containing a 1/100 dilution of protease inhibitor cocktail (Sigma) for 10 mi...
Briefly, the supernatant was incubated in 1 mL of a reaction buffer consisting of NAD (0.5 mM), triethanolamine-HCl (50 mM), MgCl2 (7.5 mM), EDTA (3.75 mM), pH 7.4. The concentration of glucose-6-phosphate was calculated from the difference between the level of NADH measured before and...
For the Ca2+ recovery experiment, cells were cultured by replacing the low Ca2+ medium restored with 1.8 mm Ca2+ for the remainder of the time course. In vitro binding assay Cells were lysed with 1 ml of lysis buffer (50 mm HEPES pH 7.5, 1% Triton X-100, 150 mm NaCl, 1...
Next, 0.25 volumes of 4× lysis buffer (1× lysis buffer: 30 mM HEPES, 50 mM KAc, 2 mM Mg(Ac)2, 1 mM EGTA, 10% glycerol, 1 mM DTT, 0.1 mM Mg-ATP, 0.5 mM Pefabloc, 10 ng/mL Leupeptin, 10 ng/mL Pepstatin A, and 0.2% v/v Triton X-100, pH 7.2...