HEK293 Whole Cell Lysate

SKU: orb348669

Description

HEK293 Whole Cell Lysate

Images & Validation

• Application Notes:
  ready-to-use lysates are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Lysates are prepared in denaturing buffer WITHOUT dissociating agents (i.e. no 2-mercaptoethanol or dithiothreitol has been added). Heat lysate 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 10-20 μl depending on the size format of your gel.

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Key Properties

• Purity: HEK293 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 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 2X SDS-PAGE sample buffer was added.
• Source: Human
• Biological Origin: Human
• Conjugation: Unconjugated

Storage & Handling

• Storage: Store vial at -70° C or COLDER. For extended storage, aliquot contents to minimize freeze/thaw cycles.
• Form/Appearance: Liquid (sterile filtered)
• 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)
• Concentration: 1.0 mg/ml
• Expiration Date: 12 months from date of receipt.
• Dry Ice Shipping: Please note: This product requires shipment on dry ice. A dry ice surcharge will apply.
• Hazard Information: Non-Toxic
• Disclaimer: For research use only

Alternative Names

HEK293 lysate, Whole Cell Lysate