Author: Su, Xiaoping; Qian, Cheng; Zhang, Qian; Hou, Jin; Gu, Yan; Han, Yanmei; Chen, Yongjian; Jiang, Minghong; Cao, Xuetao
Title: miRNomes of haematopoietic stem cells and dendritic cells identify miR-30b as a regulator of Notch1 Document date: 2013_12_6
ID: 4vo7n6nh_37
Snippet: Immunoblot. Cells were lysed using M-PER Protein Extraction Reagent (Pierce, Rockford, IL, USA) supplemented with protease inhibitor cocktail (Calbiochem). Protein concentrations of the extracts were measured with BCA assay (Pierce) and equalized with the extraction reagent. Equal amount of the extracts were loaded and subjected to SDS-PAGE, transferred onto nitrocellulose membranes. The membrane was then incubated in a 1:1,000 dilution of a prim.....
Document: Immunoblot. Cells were lysed using M-PER Protein Extraction Reagent (Pierce, Rockford, IL, USA) supplemented with protease inhibitor cocktail (Calbiochem). Protein concentrations of the extracts were measured with BCA assay (Pierce) and equalized with the extraction reagent. Equal amount of the extracts were loaded and subjected to SDS-PAGE, transferred onto nitrocellulose membranes. The membrane was then incubated in a 1:1,000 dilution of a primary antibody in 5% BSA-TBS-T. After three washes in TBS-T, the membrane was incubated with 1:5,000 dilution of the corresponding secondary antibody in 5% non-fat dry milk-TBS-T and respective proteins were then blotted. 36 .
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