Human A549 Whole Cell Lysate

• Cat.No.: A549-011HCL
• Product Overview: Ready-to-use whole cell lysates are derived from cell lines or tissues 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 and immunoblot analysis using antibodies to key cell signaling components to confirm the presence of both high molecular weight and low molecular weight proteins.

Specification

• Species: Human
• Form: Liquid (sterile filtered)
• Specificity: The cells were grown in medium supplemented with 10% fetal bovine serum. Cells were washed with PBS and then incubated on ice in modified RIPA buffer 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 2 mg/mL and then an equal volume of 2× SDS-PAGE sample buffer was added.
• Applications: WB, SDS-PAGE
• Cell Line: Human A549 (lung epithelial carcinoma)
• Lysate Fractionation: Whole Cell Lysate
• Lysate Stimulation: Not Stimulated
• Culture Type: Tissue Culture
• Induction: None (Control)
• Concentration: 1.0 mg/mL by BCA assay
• Buffer: 1× SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8)
• Preservative: None
• Stabilizer: 10% (v/v) Glycerol
• Shipping: Dry Ice
• Storage: Store vial at -70 centigrade or COLDER. For extended storage, aliquot contents to minimize freeze/thaw cycles.
• Expiration: Expiration date is three months from date of receipt.