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Catalog Number | orb348687 |
---|---|
Category | Tools |
Description | MCF-7 Cell Nuclear Extract |
Tested applications | WB |
Concentration | 1.0 mg/ml |
Dilution range | WB: User Optimized |
Form/Appearance | Liquid (sterile filtered) |
Purity | 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. |
Conjugation | Unconjugated |
Hazard Information | Non-Toxic |
Source | Human |
Storage | Store vial at -70° C or COLDER. For extended storage, aliquot contents to minimize freeze/thaw cycles. |
Buffer/Preservatives | Preservative: None. Stabilizer: 10% (v/v) Glycerol. 1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8) |
Alternative names | MCF-7 Cell Nuclear Extract, MCF7 Cell Nuclear Extr Read more... |
Note | For research use only |
Application notes | 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. |
Expiration Date | 12 months from date of receipt. |
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