Thermo Scientific M-PER Mammalian Protein Extraction Reagent was developed as an effective yet milder alternative to RIPA buffer. M-PER reagent uses a non-denaturing detergent to prepare total cell lysate that is compatible with many downstream assay...
Thermo Scientific M-PER Mammalian Protein Extraction Reagent was developed as an effective yet milder alternative to RIPA buffer. M-PER reagent uses a non-denaturing detergent to prepare total cell lysate that is compatible with many downstream assay...
GFP and GFP-ATR (4 mg of protein lysate per sample) were immunoprecipitated by incubating the lysates with 200 μl of anti-GFP-conjugated magnetic beads overnight on a rotor at 4°. Beads were washed (with Lysis buffer) and eventually pooled before elution with sample buffer. Proteins...
(二)、细胞蛋白液制备 (preparation of cell lysate) 21 (三)、蛋白质定量 (protein assay) 22 (四)、蛋白质电泳 (Western blotting) 2...cetd.tmu.edu.tw|基于122个网页 2. 细胞蛋白质制备 二、 细胞蛋白质制备(Preparation of cell lysate) 24 三、 蛋白质定量法(Protein assay) 25 四、 西方墨点法...
Cell lysates were prepared by lysing cells in ice-cold RIPA buffer (25 mM Tris–HCl pH 7.6, 0.15 M NaCl, 1% IGEPAL CA-630, 1% sodium deoxycholate and 0.1% SDS) containing 1× protease inhibitors (Thermo Scientific). Lysate was cleared by centrifugation at 20,000g for 10 min at...
buffer at 4°C. As an example, 7mL of cell lysate is loaded into the dialysis tubing (3.5kDa MWCO, SnakeSkin prewet according to the manufacturer's instructions, Thermo Scientific). The sample (in the dialysis tubing) is placed in 4L of buffer with stirring for 1.5–2h at 4°C. ...
G, USP10 was knocked out from H1299 cells by its specific sgRNA species, 48 h later, cells were treated with CHX for indicated periods, followed by cell lysate preparation and IB assay. H, the statistical analysis of PTEN stability based on G. HEK293T, human embryonic kidney 293T; IB,...
For preparation of 10 mL staining solution, we mixed 9985 μL sterile filtered 1× PBS buffer (DULBECCO, Biochrom AG), 5 μL Tween20 (ACROS Chemicals), and 50 μL propidium iodide stock solution (1 mg/mL in distilled water; Becton Dickinson). This solution was aliquoted into FACS-tubes...
(50 µg/ml) for 4 h. Briefly, the cells were washed with cold PBS and lysed with the cell lysis buffer [PIPES, KCl, mgcl2, DTT, EDTA, EGTA containing PMSF, antipain, leupeptin, and aprotinin] on ice. The cell lysates were centrifuged (12000 g for 5 min at 4 °C)...
Sample concentration.10-20 μL of cell lysates at 1x107cells per mL. (This is typically equivalent to 15-30 μg of total protein). Adjust up or down to obtain desired signal strength and low background. Blocking buffer.Start with 5% nonfat dry milk for block, and 2% nonfat dry milk ...