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 ...
For western blot analysis, cells were washed twice with PBS at 4 °C, and 0.1 ml of lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.25% sodium deoxycholate, 0.1% Nonidet P-40, 0.1% Triton X-100, 50 mM NaF, 1 mM dithiothreitol, 0.5 mM phenylmethylsulfonyl fluoride, 50 ...
The maximal respiration was measured following the injection of 1.5 mM of the uncoupling agent FCCP, then 100 nM rotenone and 1 mM antimycin were added to stop ATP-linking respiration. The cells were repeated four times per group. Western blot analysis ADSCs were lysed using RIPA lysis buffer...
After 36 h incuba- tion, the cells were exposed to hypoxic conditions for 3 h or left in normoxic condition as parallel control, and then suspended in RIPA lysis buffer. The whole of cell lysates were cultured with mouse monoclonal anti-HA (clone 16B 12; BabCO) or anti-His (Sigma...
Around 10 mg of leaf discs with 48 h of agroinfiltration were harvested and ground into fine powder in liquid nitrogen and homogenized in Passive Lysis Buffer (PLB, Promega), following with centrifugation at 13,000 rpm for 1 min. The clear supernatant was further diluted by 20 times...
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...
Cells were lysed in ALP lysis buffer [10 mM Tris-HCl (pH 7.5), 0.5 mM MgCl2, 0.1% Triton X-100] and were preincubated for 10 min at room temperature; then 100 μl of substrate solution [1 M diethanolamine buffer (pH 9.8), 0.5 mM MgCl2, 15 mM p-...
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...
and resuspended in 25 ml of lysis buffer (10 mM Tris HCl (pH 8), 0.1 M NaCl, 1.0 mM β-mercaptoethanol, 5% glycerol, 0.5 mM imidazole Triton X-100 0.02%). Cells were sonicated (Sonifier Cell DisruptorB-30; Branson Sonic Power. Co., Danbury, CT) on ice using 12 ...