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| Product Name | MCF-7 Cell Nuclear Extract |
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| Description | The cells were grown in EMEM supplemented with 10% FBS (Fetal Bovine Serum). 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 | 200 µg |
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| Concentration | 1.0 mg/mL |
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| Applications | Western Blot. Ready-to-use nuclear extracts are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Nuclear extracts are supplied in denaturing buffer without dissociating agents. Heat nuclear extract 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 10-30 ?l depending on the size format of your gel. |
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| Other Names | MCF-7 Cell Nuclear Extract, MCF7 Cell Nuclear Extract, MCF-7 Nuclear Lysate, MCF7 Lysate Nuclear Extract |
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| Gene, Accession, CAS # | n/a |
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| Catalog # | W09-001-A85 |
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| Order / More Info | MCF-7 Cell Nuclear Extract from ROCKLAND IMMUNOCHEMICALS INC. |
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| Product Specific References | n/a |
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