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| Product Name | HeLa Whole Cell Lysate Nocodozole Stimulated |
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| Description | The cells were grown in DMEM supplemented with 10% FBS (Fetal Bovine Serum). Cells were treated with 3 µg/ml nocodazole for 1 h. Cells were washed in PBS and incubated on ice in modified RIPA buffer containing 150 mM sodium chloride, 50 mM Tris Cl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.5% sodium deoxycholic acid and 0.1% SDS to lyse the cells. Protein integrity is 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 and 1 µM Pepstatin A). Phosphatase inhibitors sodium fluoride, sodium orthovanadate, sodium pyrophosphate and ?-glycerophosphate were also added. Cell debris was removed by centrifugation. Protein concentration was determined by Lowry assay using a commercially available kit. The 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 Whole Cell Lysate Nocodozole Stimulated, HeLa Nocodozole Stimulated Whole Cell Lysate, HeLa Nocodozole Stimulated WCL, HeLa WCL Nocodozole Stimulated |
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| Gene, Accession, CAS # | n/a |
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| Catalog # | W09-001-A81 |
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| Price | |
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| Order / More Info | HeLa Whole Cell Lysate Nocodozole Stimulated from ROCKLAND IMMUNOCHEMICALS INC. |
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| Product Specific References | n/a |
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