Tissue preparationAfter deep anesth

Tissue preparation
After deep anesthesia with sodium pentobarbital (60 mg/kg), Ntg, mtauKO, 3xTg-AD and 3xTg-AD/mtauKO mice were perfused trans- cardially with 0.1 M phosphate-buffered saline (PBS), pH 7.4. Half of the brain was fixed for 48 h in 4% paraformaldehyde in 0.1 M phos- phate buffer saline, pH 7.4 and cryoprotected in 30% sucrose for immu- nohistochemical analysis, whereas the other half was frozen in dry ice for biochemical analysis. Thick (40 μm) free-floating sections were obtained using a freezing microtome (Leica SM 2010 R) and serially col- lected (each series contained sections that represented 1/7th of the total brain) in cold PBS and 0.02% sodium azide.
Protein extracts were prepared by homogenizing whole brain hemi- sphere samples in T-per (Thermo Fisher Scientific, Rockford, IL) extrac- tion buffer (150 mg/ml), complemented with a protease inhibitor (Complete Mini Protease Inhibitor Tablets, Roche Diagnostics GmbH, Germany) and phosphatase inhibitor (5 mmol/l, Sigma-Aldrich, St. Louis, MO), followed by centrifugation at 100,000 ×g for 1 h. Protein concentration in the supernatant was determined using the Bradford assay.

ผลลัพธ์ (อังกฤษ) 1:

tissue preparation
after deep anesthesia with sodium pentobarbital (60 mg / kg), ntg, mtauko, 3xtg-ad and 3xtg-ad/mtauko mice were perfused trans-cardially with 0.1 m phosphate-buffered saline (pbs), ph 7.4. half of the brain was fixed for 48 h in 4% paraformaldehyde in 0.1 m phos-phate buffer saline, ph 7.4 and cryoprotected in 30% sucrose for immu-nohistochemical analysis,.whereas the other half was frozen in dry ice for biochemical analysis. thick (40 μm) free-floating sections were obtained using a freezing microtome (leica sm 2010 r) and serially col-lected (each series contained sections that represented 1/7th of the total brain) in cold pbs and 0.02% sodium azide.
protein extracts were prepared by homogenizing whole brain hemi-sphere samples in t-per (thermo fisher scientific, rockford, il) extrac-tion buffer (150 mg / ml), complemented with a protease inhibitor (complete mini protease inhibitor tablets, roche diagnostics. gmbh, germany) and phosphatase inhibitor (5 mmol / l, sigma-aldrich, st. louis, mo), followed by centrifugation at 100,000 × g for 1 h.

ผลลัพธ์ (อังกฤษ) 3:

After Tissue preparation deep anesthesia with sodium pentobarbital (60 mg/kg), Ntg, mtauKO, 3 xTg - AD and 3 xTg - AD/mtauKO mice were perfused trans-cardially M 0.1 with phosphate-buffered saline (PBS), pH 7.4. Half of the brain was fixed for 48 h in 4% paraformaldehyde in 0.1 M phos- phate buffer saline, 7.4 pH and cryoprotected in 30% sucrose for immu-nohistochemical analysis,whereas the other half was frozen in dry ice for biochemical analysis. Thick (40 µm) free-floating sections were obtained using a freezing microtome (Leica SM 2010 R), and serially col-lected (each series contained sections that represented 1 /7 th of the total brain) in cold PBS and 0.02 % sodium azide.
Protein extracts were prepared by homogenizing whole brain hemi-sphere samples in T-per (Thermo Fisher Scientific, Rockford, IL) extrac-tion buffer (150 mg/ml), complemented with a protease inhibitor (Complete Mini Protease Inhibitor Tablets, Roche Diagnostics GmbH, Germany), and phosphatase inhibitor (5 mmol/L, Sigma - Aldrich, St.Louis, MO), followed by centrifugation at 100,000 G for 1 H × .Protein concentration in the supernatant was determined using the Bradford assay.