Sentences with phrase «in lysis buffer»

Hippocampus or frontal cortex samples were homogenized in lysis buffer (10 mm Tris, pH 7.5, 10 mm NaCl, 3 mm MgCl2, 1 mm EDTA, and 0.05 % NP - 40) with protease inhibitors (Halt protease inhibitor cocktail, Thermo Scientific) and protein concentration was determined by Bradford assay (Coomassie Plus, Thermo Scientific).

After an overnight incubation at 4 ° C, for thorough stabilization, samples were homogenized in lysis buffer and total RNA was isolated using the GE Illustra RNAspin Isolation Kit (GE Healthcare) according to the manufacturer's protocol.

At the end of the desired treatment times, cell lysates were prepared in lysis buffer (1 % NP - 40, 0.5 mM Tris - HCl (pH 7.5), 0.14 M NaCl, 5 mM KCl, 5 mM EDTA, and 1 mM phenylmethylsulfonyl fluoride) plus serine / threonine phosphatase inhibitor cocktail.

Total proteins were harvested in lysis buffer (20 mM Tris - HCl [pH 8.0], 150 mM NaCl, phosphatase inhibitors [Pierce], protease inhibitors [C complete protease inhibitor cocktail, Roche], and 1 % Triton X-100).

Not exact matches

At 48 hours, cells were washed twice with PBS and lysed in 1 × passive lysis buffer (Dual - Luciferase Reporter Assay, Promega).

The SVCs were resuspended in erythrocyte lysis buffer and incubated at room temperature for 5 minutes.

Single animals were placed into 5 µl of worm lysis buffer (10 mM Tris — HCl pH 8.3, 50 mM KCl, 2.5 mM MgCl2, 0.45 % NP - 40, 0.45 % Tween 20, 0.01 % Gelatin and 500 µg / ml fresh proteinase K) in a PCR tube.

Spleens were processed into cell suspensions through a 70 - μm strainer by a syringe plunger in RPMI 1640 with 10 % FBS and then treated with ACK lysis buffer to remove RBCs.

Reactions were terminated with 1 ml of ice - cold PBS, and cells were centrifuged and resuspended in 50 μl of cell lysis buffer containing 1 % NP - 40 and protease inhibitors.

Cells were lysed in RIPA lysis buffer.

Hippocampal and entorhinal cortex tissue samples were homogenized in 10 volumes of RIPA lysis buffer (50 mM Tris, pH 7.5, 150 mM NaCl, 0.1 % sodium dodecyl sulfate and 0.5 % deoxycholate, and 1 % NP40) containing a cocktail of protease and phosphatase inhibitors (20 mg / ml each of pepstatin A, aprotinin, phosphoramidon, and leupeptin; 0.5 mM 4 -(2 - aminoethyl) benzenesulfonyl fluoride hydrochloride; 1 mM EGTA; 5 mM fenvalerate; and 5 mM cantharidin).

NHLF cells transfected with siRNA molecules were lysed in High Salt ELB lysis buffer [1 M Tris pH 8.0, 1 % NP - 40, 250 mM NaCl, 5 mM EDTA] supplemented with protease and phosphatase inhibitors (1x G - Biosciences Protease Arrest, 200 µM Na3VO4, and 1 mM PMSF).

Cell pellets were resuspended in 1 ml cell lysis buffer [5 mM Pipes pH 8.0, 85 mM KCl, 0.5 % NP - 40] containing protease inhibitors and incubated for 10 min on ice.

For this protocol, samples were lysed in 1 × RIPK2 lysis buffer [50 mM Tris (pH 7.5), 0 mM MgCl2, 1 % Triton X-100, 1 mM dithiothreitol, 1 mM EDTA, 1 mM EGTA, with freshly added 1 mM β - glycerophosphate and protease inhibitors (phenylmethylsulfonyl fluoride and aprotinin)-RSB- and immunoprecipitated overnight using 1.5 μg rabbit anti-RIPK2 antibody.

Confluent 10 cm plates were fixed with 1 % PFA at room temperature for 10 minutes, lysed in SDS lysis buffer (50 mM Tris, 10 mM EDTA, 1 % SDS, Roche Complete protease inhibitors, pH 8.1), scraped and collected into 1.5 mL microcentrifuge tubes, and DNA was sonicated to 200 — 800 bp fragments with a Branson Sonifier 250 set to 30 % power / 90 % duty, four 10 second pulses.

Each single cell was carried through two rinses of PBS using a finely drawn glass capillary pipette under a dissecting microscope and then transferred in a minimal volume (0.5 µL) to a PCR tube containing lysis buffer, protease, tRNA and poly - T gripNA ™ probe (mTRAP ™ midi - kit, Active Motif, cat.

Tubes were spun, the nuclear lysis buffer with proteinase inhibitor was added and sonicated to form shared chromatin of about 200 — 1000 base pairs in length (data not shown).