Selected article for: "blot analysis and HeLa cell"
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.

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    • lysis buffer and Pierce Assay kit: 1, 2
    • lysis buffer and primary antibody: 1, 2, 3, 4, 5, 6, 7, 8, 9
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    • lysis buffer and Protein Pierce Assay kit: 1
    • lysis buffer and SIGMA protease: 1, 2, 3, 4, 5, 6, 7
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    • lysis buffer ml and protein concentration: 1, 2, 3, 4
    • lysis buffer ml and protein extraction: 1
    • lysis buffer ml and SIGMA protease: 1, 2
    • mammalian cell and primary antibody: 1
    • mammalian cell and protein concentration: 1
    • mammalian cell and western blot analysis: 1, 2
    • PBS wash and primary antibody: 1, 2, 3, 4
    • PBS wash and protein concentration: 1, 2
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