MDA-MB-231 Whole Cell Lysate

SDS-Page: MDA-MB-231 Whole Cell Lysate [NBP3-18060] - SDS PAGE Results of MDA-MB-231 Whole Cell Lysate. Lane 1: MDA-MB-231 Whole Cell Lysate Reduced [10ug]. Lane 2: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 3: MDA-MB-231 Whole Cell Lysate Non-Reduced [10ug]. 4-20% Gel, Coomassie Stained.

• Reactivity: Hu
• Applications: WB, IP, PAGE, ChIP

MDA-MB-231 Whole Cell Lysate Summary

• Preparation Method: 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 uM Aprotinin, 5 uM Bestatin, 1.5 uM E-64, 2 uM Leupeptin Hemisulfate, 1 uM 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 with 1X RIPA buffer.
• Purity: N/A

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP)
  • Immunoprecipitation
  • SDS-Page
  • Western Blot
• Application Notes: This product has been tested by SDS-PAGE. Multi-purpose MDA-MB-231 Whole Cell Lysate 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 95C for 5 minutes. Sample is ready for use in immunoprecipitation and ELISA experiments, conditions should be optimized by the user. It is recommended to use TrueBlot IP reagents for immunoprecipitation experiments.

Packaging, Storage & Formulations

• Storage: Store at -70C. Avoid freeze-thaw cycles.
• Buffer: 1X RIPA Buffer with HALT Protease and Phosphatase Inhibitors.
• Preservative: No Preservative
• Purity: N/A

Lysate Details for Array

• Type: Cell
• Subcellular Fraction: Whole

Background

Multi-purpose MDA-MB-231 Whole Cell Lysates are derived from cell lines using highly refined extraction protocols to ensure exceptionally high quality, protein integrity and lot-to-lot reproducibility. All extracts are tested by SDS-PAGE using 4-20% gradient gels.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.