| 產(chǎn)品詳情 |
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| Product Name | A431 Whole Cell Lysate EGF Stimulated |
| Description | The cells were grown in Dulbecco's medium supplemented with 10% fetal bovine serum. 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. |
| Size | 500 μg |
| Concentration | 1mg/mL |
| Applications | Western Blot. A431 Whole Cell Lysate - EGF Stimulated 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. |
| Other Names | Cell Lysate, Stimulated Lysate, A431 Lysate EGF Stimulated, A431 Lysate Epidermal growth factor Stimulated |
| Gene, Accession, CAS # | n/a |
| Catalog # | W09-000-362 |
| Price | |
| Order / More Info | A431 Whole Cell Lysate EGF Stimulated from ROCKLAND IMMUNOCHEMICALS INC. |
| Product Specific References | n/a |
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