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Catalog Number | orb348701 |
---|---|
Category | Tools |
Description | MDA-MB-231 Cell Nuclear Extract |
Tested applications | ChIP, IP, WB |
Concentration | 4.0 mg/ml |
Dilution range | ChIP: User Optimized, IP: User Optimized, WB: User Optimized |
Form/Appearance | Liquid (sterile filtered) |
Purity | MDA-MB-231 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 4 mg/ml in RIPA buffer containing protease and phosphatase inhibitors. |
Conjugation | Unconjugated |
Hazard Information | Non-Toxic |
Source | Human |
Storage | Store vial at -70° C or COLDER. For extended storage, aliquot Nuclear Extract to minimize freeze/thaw cycles. |
Buffer/Preservatives | Preservative: None. Stabilizer: None. 1X RIPA Buffer with HALT Protease and Phosphatase Inhibitors |
Alternative names | MDA-MB-231 lysate nuclear extract, cell lysate, MD Read more... |
Note | For research use only |
Application notes | Multi-purpose MDA-MB-231 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. |
Expiration Date | 12 months from date of receipt. |
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