For the first-step immunoprecipitation assay, whole-cell extracts were prepared by using low-salt buffer supplemented with a 5 mg/ml protease inhibitor cocktail (Roche). Lysates were incubated with the anti-FLAG beads overnight. The immunoprecipitates were washed 3–5 times with low-salt ...
The infection of cells was performed as described by Stubenrauch and Pfister (1994); 72 h postinfection cells were resuspended in a total volume of 10 ml of PBS containing 1 mM phenylmethylsulfonyl fluoride (PMSF) and protease inhibitor cocktail (Roche, Mannheim). The suspension was frozen ...
Samples were homogenized in TNE buffer containing 0.5% Triton X-100 (Sigma), 5 mM NaF, 100 μM Na3VO4 and a cocktail of protease inhibitors (CompleteTM, Roche) and briefly sonicated. Cell debris and nuclei were pelleted by centrifugation (800 g, 30 min at 4 °C). Protein...
The membranes are then cut out and transferred to scintillation vials followed by the addition of 1 mL of H2O and 6 mL of Ready Safe™ liquid scintillation cocktail to each vial. The vials are counted on a Beckman LS 6500 scintillation counter....
Since a cytomorphologic and histologic classification of thymomas seems not to be a useful prognosticator in animals, the use of a pan-specific cocktail of antibodies for cytokeratins is sufficient for the diagnosis of these tumors in veterinary medicine. Thymoma should be differentiated from other ...
Briefly, one gram of the corn flour CRM was gravimetrically fortified with 0.5 g of the cocktail internal standard solution (approximately 1:1 isotope ratio). Samples were extracted with pure water (10 mL) for 30 min with constant shaking to achieve adequate equilibrium of the analytes and the...
Protein extracts for Western blots were isolated from LCLs by homogenization in radio-immunoprecipitation assay (RIPA) buffer containing 1 mM phenylmethylsulfonyl fluoride (PMSF; Sigma-Aldrich) and 1 protease inhibitor cocktail tablet per 10 ml RIPA buffer (Complete Mini; Roche Diagnostics, Mannheim, ...
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Type 2 diabetes (T2D) is a chronic metabolic condition associated with obesity, oxidative stress-mediated inflammation, apoptosis, and impaired insulin signaling. The utilization of phytochemical therapy generated from plants has emerged as a promising a
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