Sentence examples for fractionation buffer from inspiring English sources

The supernatant was collected (cytosolic fraction) and the pellet resuspended in the Fractionation Buffer Mix (mitochondrial fraction).

The supernatant was collected (cytosolic fraction) and the pellet was suspended in a fractionation buffer mix (mitochondrial fraction).

The pellet (nuclear fraction) was washed twice in fractionation buffer, before addition of SDS-Urea loading buffer (8 M Urea, 2% SDS, 50 mM Tris HCl [pH 6.8], 0.3% bromophenol blue).

Cytoplasmic and nuclear protein fractions were extracted in subcellular fractionation buffer (150 mM NaCl, 50 mM HEPES pH 7.4, 0.5% NP40, 2 mM PMSF and Protease Inhibitor Cockail) alternatively as reported by Joseph et al. Total protein extracts were made directly in Laemmli buffer.

Cells were transfected using Polyfect (Qiagen) and fractionated 48 hours after transfection in fractionation buffer [10 mM Tris pH 7.5, 2.5 mM MgCl2, 10 mM DTT, PhosStop (phosphatase inhibitor cocktail; Roche), complete (protease inhibitor cocktail; Roche), 80 µg/ml Digitonin] as described (see Western blot).

The integration assay was performed as follows: Post-nuclear supernatant (800 µg total protein) or the mitochondria-enriched fraction P1 (150 µg total protein) in cell fractionation buffer were incubated with in vitro-synthesized proteins for 60 minutes at 4°C.

Cycloheximide was added to a final concentration of 0.1 mg/ml, and cells were harvested (4000 rpm, 4 min, 4°C) and resuspended in 400 µl of ice-cold membrane-fractionation buffer (20 mM Hepes-KOH pH 7.6, 100 mM potassium acetate, 5 mM magnesium acetate, 2 mM dithiothreitol, 0.1 mg/ml cycloheximide and 0.1 mM PMSF) either with or without additional 20 mM EDTA.

Subcellular fractionation with extraction buffer containing between 0.1 and 0.8 M NaCl, demonstrated that wild-type GATA3 was extracted efficiently from nuclei at moderate salt concentration (0.4 M NaCl) from nuclei in T47D cells.

For fractionation of the NE, a buffer (buffer H) containing 50 mM HEPES-NaOH (pH 7.9), 20% (v/v) glycerol, and 1 mM DTT in the presence of the appropriate concentrations of KCl was used for ion-exchange chromatography for purification of IREF-2.

To validate the activity of miR-21 on MMP inhibition, MMP2 inhibitor I (Merck) was either added at a final concentration of 25  μM to CM/CMSCC for 1 h before fractionation or to developing buffer following fractionation.

A second cell pellet obtained at the same time was homogenized in ice-cold cell disruption buffer without fractionation to give a whole cell lysate.