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...
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...
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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...
Cells were lysed in ice-cold NP40 lysis buffer [150 mM NaCl, 1% IGEPAL CA-630, 50 mM Tris-HCl (pH 8.0)] containing protease inhibitors (cocktail set III; Calbiochem, Merck Millipore) and phosphatase inhibitors (Halt phosphatase inhibitor cocktail; Thermo Scientific, USA). Samples ...
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...
The filtered solution was centrifuged at 110×g for 7 min, 4 °C, and the chloroplast pellet was re-suspended in 1.6 mL chloroplast lysis buffer (62.5 mM Tris–HCl, pH 5.7, 2 mM MgCl2). After incubating on ice for 15 min with gentle vortexing at 5-min intervals, the supernatants (...
17 Cell or tissue samples were lysed in RIPA lysis buffer (25 mmol/L Tris-HCl, pH 7.4, 150 mmol/L NaCl, 1% NP-40, 0.1% sodium dodecyl sulfate, 0.5% sodium deoxycholate) supplemented with protease inhibitor cocktails (Roche) by a 30-minute incubation at 4°C. The protein concentration ...
Cells were collected, lysed and sonicated at 4 °C in a lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.5% SDS, 5 mM sodium pyrophosphate, protease and phosphatase inhibitor tablet). Cell lysates were centrifuged at 13,000g for 15 min, and the supernatant was recove...