| 產品詳情 |
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| Product Name | HL-60 Cell Nuclear Extract |
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| Description | HL-60 cells were grown in Ioscove'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 4 mg/ml in RIPA buffer containing protease and phosphatase inhibitors. |
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| Size | 200 μg |
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| Concentration | 4.0 mg/mL |
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| Applications | Chip, Immunoprecipitation, Western Blot. Multi-purpose HL-60 nuclear extracts are especially prepared as positive control for multiple assays including western blot, immunoprecipitation (IP), capture ELISA or other assays requiring native protein sample. For separation by SDS-PAGE and subsequent western blot analysis, lysates should be diluted by user to desired concentration in SDS-PAGE buffer with 2-mercaptoethanol or dithiothreitol as the reducing agent and heated to 95° C for 5 minutes. Sample is ready for use in immunoprecipitation and ELISA experiments, conditions should be optimized by the user. Rockland recommends its Trueblot IP reagents for immunoprecipitation experiments. |
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| Other Names | HL-60 lysate nuclear extract, HL60 nuclear lysate, HL-60 Nuclear lysate |
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| Gene, Accession, CAS # | n/a |
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| Catalog # | W09-001-GK9 |
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| Price | |
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| Order / More Info | HL-60 Cell Nuclear Extract from ROCKLAND IMMUNOCHEMICALS INC. |
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| Product Specific References | n/a |
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