1. The buffer system The pH of the solution is critical. Proteins may precipitate or become unstable when the pH is outside of the physiological range. To avoid this situation, a buffer system such as Tris-HCl is recommended. Besides buffering solutions in this range, a Tris-HCl buffer pre...
How many mL of 4.4 M HCl solution must be added to 3.1 L of a 0.20 M K2HPO4 solution to prepare a buffer with a pH of 7.43? How many milliliters of 4.2 M HCl must be added to 3.1 L of 0.16 M K2HPO4 to prepare a...
Cells were lysed in RIPA buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.1% sodium deoxycholate, 0.5% SDS) and immunoprecipitation carried out using the Dynabeads Protein G Immunoprecipitation Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. Elut...
For identifying the interacting partners of TM9SF4, A2780 and NIH3T3 cells, with or without GFP-TM9SF4 overexpression, were lysed in lysis buffer (50 mM Tris-HCl, pH 7.5; 150 Mm NaCl; 1% NP-40; 0.5% sodium deoxycholate; 0.2% Triton X-100 and protease inhibitor cocktail) at 4 ...
Two days after transfection, the cells were harvested, washed with PBS, and treated with 100 μl of lysis buffer (0.25 M Tris-HCl, pH 7.8 + 0.1% NP40) for 30 min at 4°C and cellular debris was removed by centrifugation for 10 min. For the β-galactosidase assay, 30 μl of cell...
The culture was incubated for 20h at 18C, bac- teria were harvested by centrifugation (6000 g, 10 min, 4C) and resuspended in lysis buffer (50 mM Tris/HCl at pH 8, 150 mM NaCl, 2 mM b-mercaptoethanol). After sonication and re...
HCLUSCRYPTPROVIDER structure (Windows) C-C++ Code Example: Reading Messages Asynchronously Using Completion Ports C-C++ Code Example: Creating a Security Descriptor FaultHandlerActivity.System.Workflow.ComponentModel.IActivityEventListener<System.Workflow.ComponentModel.ActivityExecutionStatusChangedEventArgs>.OnEve...
(Life Technologies) 1:4000 in a hypotonic lysis buffer (10 mM Tris HCl (pH 8), 5 mM EDTA, 0.1 % Triton X-100) overnight in the dark at 4 °C and then quantified by fluorimetry, measured using a FLUOstar Optima (BMG LabTech, Ortenberg, Germany) set to 485 nm excitation and 535...
Fully differentiated adipocyte monolayers were rinsed with ice-cold PBS and homogenised in a lysis buffer containing 50 mM Tris-HCl pH 7.4, 100 mM NaCl, 50 mM NaF, 5 mM EDTA, 2 mM EGTA, 40 mM beta-glycerophosphate, 10 mM sodium pyrophosphate, 1% Triton X-100 and protease...
To isolate the DNA, digit tissue was treated with Proteinase K (20 µg/ml) in 500 µL of Tris-HCl buffer (Composition: 100 mM Tris-HCl (pH8.0); 5 mM EDTA; 200 mM NaCl; 0.2% SDS) overnight at 56°C. The DNA was precipitated by adding equal volume of isopropanol ...