LNCaP Cell Nuclear Extract

• Catalog Number: orb348695
• Category: Tools
• Description: LNCaP 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: LNCap cells were grown in RPMI-1640 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: LNCaP lysate nuclear extract, cell lysate, LNCaP N
• Note: For research use only
• Application notes: Multi-purpose LNCaP 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.