In these experiments, a mutated protein (400 nM) was added to a stirred cuvette containing D-CaM (12.5 nM) in Fluorescence buffer (in the presence or absence of Ca2+) and after two minutes, another mutant (400 nM) was added.
SRL showed good solubility and stability at pH 4.0 (both in acetate buffer as well in phosphate buffer) in the presence of block copolymers.
Cryogrinded powders from each vial were resuspended in 2 ml RA1 lysis buffer in the presence of 1% β-mercaptoethanol, then pooled together and transferred to a single NucleoSpin Filter Midi.
Linezolid reacted with palladium to form 1 1 binary cationic chelate which further reacted with eosin dye to form 1 1 ternary ion association complex at pH 4 of Walpole's acetate buffer in the presence of methyl cellulose.
This method utilizes glycerol phosphorylation to generate a product easily quantifiable at 570 nm. 2 × 106 cells were lysed in 100 μL ATP Assay Buffer in the presence of zymolyase (0.1 U/μL) and incubated for 1 hr at 37 °C.
Reactions were performed in sodium phosphate/citrate buffer in the presence of 2 mM H2O2 at 25��C.
For control heat treatment experiments (without addition of calcium), the enzyme solution was dialyzed against 0.1 M Tris-HCl buffer pH 7.5 and heating was performed in this buffer in the presence of 2 mM ethylenediaminetetraacetic acid (EDTA).
The solution was then diluted to 500 mL and standardized with lead nitrate using a pH 5.3 buffer in the presence of hydrogen peroxide by back titrating excess EDTA using xylenol orange indicator.
Protein was extracted by NET-NP40 buffer in the presence of Complete™ Protease Inhibitor Cocktail (Roche).
The eluted proteins were then incorporated into liposomes by dialysis against glycerol-free buffer in the presence of lipids.
The beads were then washed three times in 0.5 ml of resuspension buffer in the presence of 0.1 mM Mg2Cl.
