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| Product Name | Mouse Embryonic Fibroblast Whole Cell Lysate |
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| Description | The cells were grown in RPMI supplemented with 10% FBS (Fetal Bovine Serum). The lysate was prepared by first washing the cells in PBS. Washed cells are then 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, 0.25% sodium deoxycholic acid 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 include 1 mM Sodium Fluoride and 1mM Na3VO4. Cell debris was removed by membrane filtration. 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 | 250 µg |
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| Concentration | 1.0 mg/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 | Mouse Embryonic Fibroblast Whole Cell Lysate, Mouse Embryonic Fibroblast WCL, MEF Whole Cell Lysate, MEF WCL, MEF Lysate, Mouse Embryonic Fibroblast Lysate |
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| Gene, Accession, CAS # | n/a |
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| Catalog # | W10-001-371 |
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| Order / More Info | Mouse Embryonic Fibroblast Whole Cell Lysate from ROCKLAND IMMUNOCHEMICALS INC. |
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| Product Specific References | n/a |
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