Human Nocodazole Stimulated HeLa Whole Cell Lysate

• Cat.No.: HeLa-026HCL
• 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 DMEM supplemented with 10% FBS (Fetal Bovine Serum). Cells were treated with 3 μg/mL nocodazole for 1 h. Cells were washed in PBS and incubated on ice in modified RIPA buffer 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 2× SDS-PAGE sample buffer was added.
• Applications: SDS-PAGE
• Cell Line: HeLa - Human epidermoid carcinoma
• Lysate Fractionation: Whole Cell Lysate
• Lysate Stimulation: Nocodozole
• Culture Type: Tissue Culture
• Induction: Nocodazole (3 μg/mL)
• Concentration: 1 mg/mL by modified Lowry 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.