Selected article for: "Assay kit and RIPA buffer"
Author: Dong, Xiaoxv; Ni, Boran; Fu, Jing; Yin, Xingbin; You, Longtai; Leng, Xin; Liang, Xiao; Ni, Jian
Title: Emodin induces apoptosis in human hepatocellular carcinoma HepaRG cells via the mitochondrial caspase-dependent pathway
  • Document date: 2018_8_2
    • ID: 02r8i5n1_12
    Snippet: Western blot analysis. HepaRG cells were plated in 6-well plates and treated as described above. The cells were harvested, lysed with RIPA buffer for 30 min on ice. Lysates were centrifuged at 12,000 rpm for 10 min at 4˚C. Total protein concentration was determined using the bicinchoninic acid assay (BCA) protein assay kit. In a parallel experiment, cytosolic and mitochondrial extracts were prepared using the ProteoExtract ® Cytosol/Mitochondri.....
    Document: Western blot analysis. HepaRG cells were plated in 6-well plates and treated as described above. The cells were harvested, lysed with RIPA buffer for 30 min on ice. Lysates were centrifuged at 12,000 rpm for 10 min at 4˚C. Total protein concentration was determined using the bicinchoninic acid assay (BCA) protein assay kit. In a parallel experiment, cytosolic and mitochondrial extracts were prepared using the ProteoExtract ® Cytosol/Mitochondria Fractionation Kit (Millipore, Billerica, MA, USA) following the manufacturer's protocol. An equal amount of proteins was separated by electrophoresis on SDS-polyacrylamide gels (SDS-PAGE) and transferred onto polyvinylidene fluoride (PVDF) membranes. The membranes were blocked with TBST buffer containing 5% skim milk for 1 h, and then incubated with specific primary antibodies overnight at 4˚C. After three times washing with TBST buffer, the membranes were further incubated with the horseradish peroxidase-conjugated secondary antibodies (cat. no. 3700; Cell Signaling Technology) at room temperature for 1 h (37, 38) . An enhanced ECL detection system was used for visualization of target proteins (iNtRON Biotechnology, Seongnam, Korea).

    Search related documents:
    Co phrase search for related documents
    • SDS polyacrylamide gel and skim milk: 1, 2, 3, 4
    • SDS polyacrylamide gel and target protein: 1
    • SDS polyacrylamide gel and total protein: 1, 2, 3, 4, 5
    • secondary antibody and skim milk: 1, 2, 3, 4
    • secondary antibody and specific primary antibody: 1, 2, 3, 4, 5, 6
    • secondary antibody and target protein: 1
    • secondary antibody and TBST buffer: 1
    • secondary antibody and total protein: 1, 2, 3, 4, 5, 6, 7, 8, 9
    • secondary antibody and total protein concentration: 1, 2, 3
    • secondary antibody and western blot analysis: 1, 2, 3, 4, 5, 6, 7
    • skim milk and target protein: 1
    • skim milk and total protein: 1, 2, 3
    • skim milk and total protein concentration: 1
    • skim milk and western blot analysis: 1, 2
    • target protein and total protein: 1, 2, 3, 4, 5, 6, 7
    • target protein and western blot analysis: 1, 2, 3, 4, 5, 6
    • TBST buffer and total protein: 1
    • total protein and western blot analysis: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
    • total protein concentration and western blot analysis: 1