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| Product Name | HeLa Whole Cell Lysate MG-132 Treated |
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| Description | HeLa cells were grown in Dulbecco's medium supplemented with 10% fetal bovine serum. Cells were treated O/N with 0.2µg/mL of MG-132 in order to reduce degradation of ubiquitin-conjugated proteins by the proteasome. Cells were washed with PBS and then incubated on ice in modified RIPA buffer, containing 150 mM sodium chloride, 50 mM Tris HCl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.5% sodium deoxycholic acid , 0.1% SDS and 0.01% (w/v) sodium azide to lyse the cells. Protein integrity was ensured using a cocktail of protease inhibitors with broad specificity for the inhibition of aspartic, cysteine, and serine proteases as well as aminopeptidases (0.1 mM AEBSF HCl, 0.08 µM Aprotinin, 5 µM Bestatin, 1.5 µM E-64, 2 µM Leupeptin Hemisulfate, 1 µM Pepstatin A). Phosphatase inhibitors 1 mM NaF and 1 mM Na3VO4 were also added. Cell debris was removed by centrifugation. Protein concentration was determined by a modified Lowry assay using a commercially available kit. Protein concentration was adjusted to 2 mg/ml and then an equal volume of 2X SDS-PAGE sample buffer was added. |
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| Size | 500 µg |
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| Concentration | 1mg/mL |
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| Applications | Western Blot. Ready-to-use lysates are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Lysates are prepared in denaturing buffer WITHOUT dissociating agents (i.e. no 2-mercaptoethanol or dithiothreitol has been added). Heat lysate to 95° C for 5 minutes and rapidly cool. If dissociating conditions are desired, add reducing agent prior to heating. The recommended loading volume per lane is 10-20 µl depending on the size format of your gel. |
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| Other Names | HeLa Lysate MG132 stimulated, Cell Lysate, MG-132 Stimulated Lysate HeLa cell, HeLa lysate MG-132 treated |
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| Gene, Accession, CAS # | n/a |
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| Catalog # | W09-001-GV9 |
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| Price | |
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| Order / More Info | HeLa Whole Cell Lysate MG-132 Treated from ROCKLAND IMMUNOCHEMICALS INC. |
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| Product Specific References | n/a |
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