Selected article for: "HeLa cell and western blot"
Author: de Melo, Ivan S.; Jimenez-Nuñez, Maria D.; Iglesias, Concepción; Campos-Caro, Antonio; Moreno-Sanchez, David; Ruiz, Felix A.; Bolívar, Jorge
Title: NOA36 Protein Contains a Highly Conserved Nucleolar Localization Signal Capable of Directing Functional Proteins to the Nucleolus, in Mammalian Cells
  • Document date: 2013_3_13
    • ID: 0jx6mwiw_20
    Snippet: For cellular protein extraction, 1610 6 transfected HeLa cells were washed with PBS, and cell pellets were resuspended in 0.5 ml of lysis buffer (50 mM Tris-Cl, pH 7.5, 1% NP-40, 1% Nadeoxycholate, 0.1% SDS, 2 mM EDTA, 0.5 M NaCl, and 1/500 of SIGMA-protease inhibitor cocktail for mammalian cells). Preparations were incubated for 5 min in ice and then were passed through a narrow gauge needle (21G) to shear DNA. Then the lysates were cleared by c.....
    Document: For cellular protein extraction, 1610 6 transfected HeLa cells were washed with PBS, and cell pellets were resuspended in 0.5 ml of lysis buffer (50 mM Tris-Cl, pH 7.5, 1% NP-40, 1% Nadeoxycholate, 0.1% SDS, 2 mM EDTA, 0.5 M NaCl, and 1/500 of SIGMA-protease inhibitor cocktail for mammalian cells). Preparations were incubated for 5 min in ice and then were passed through a narrow gauge needle (21G) to shear DNA. Then the lysates were cleared by centrifugation (14.000 g, 10 min at 4uC). Supernatants were used as fractions with the extracted proteins. MicroBCA Protein Assay Kit (Pierce) was used to determine protein concentrations. Western blot analysis was performed as described [36] , but using 60 mg aliquots of extracted proteins and specific anti-Flag (SIGMA) or anti-eGFP (Abcam) primary antibodies.

    Search related documents:
    Co phrase search for related documents
    • Abcam primary antibody and primary antibody: 1, 2, 3, 4, 5, 6, 7
    • Abcam primary antibody and protein concentration: 1, 2
    • Assay kit and lysis buffer: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10
    • Assay kit and lysis buffer ml: 1, 2
    • Assay kit and PBS wash: 1
    • Assay kit and Pierce Assay kit: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10
    • Assay kit and primary antibody: 1, 2, 3
    • Assay kit and protein concentration: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17
    • blot analysis and lysis buffer: 1, 2, 3, 4, 5, 6, 7, 8, 9
    • blot analysis and mammalian cell: 1, 2, 3
    • blot analysis and primary antibody: 1, 2, 3, 4, 5, 6
    • blot analysis and protein concentration: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11
    • cell pellet and lysis buffer: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10
    • cell pellet and lysis buffer ml: 1, 2, 3
    • cell pellet and Pierce Assay kit: 1
    • cell pellet and protein concentration: 1, 2, 3, 4
    • extract protein and protein concentration: 1, 2
    • gauge needle and lysis buffer: 1
    • gauge needle and protein concentration: 1, 2