Mouse Embryonic Fibroblast Whole Cell Lysate

• Cat.No.: Embryonic Fibroblast-070MCL
• 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: Mouse
• Form: Liquid (sterile filtered)
• Specificity: The cells were grown in RPMI supplemented with 10% FBS (Fetal Bovine Serum). The lysate was prepared by first washing the cells in PBS. Washed cells are then 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 include 1 mM Sodium Fluoride and 1mM Na3VO4. Cell debris was removed by membrane filtration. 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: Mouse Embryonic Fibroblast
• Lysate Fractionation: Whole Cell Lysate
• Lysate Stimulation: Not Stimulated
• Culture Type: Tissue Culture
• Induction: None (Control)
• Concentration: 1.0 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.