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MCF-7 Whole Cell Lysate Lysate

WB Whole Cell Lysate Cell Lysate MCF7 Cells Reactivité: Humain
N° du produit ABIN964020
$244.50
Plus frais de port $45.00 et $24.00 pour carboglace
500 μg
local_shipping Destination: Etats-Unis
Envoi sous 1 à 2 jours ouvrables
  • Espèce de la protéine
    Humain
    Espèce du lysat
    Human Cells
    Application
    Western Blotting (WB)
    Specificité
    The 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 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.
    Attributs du produit
    Cell Line: MCF7 - Human Epithelial (Mammary)
    Induction: None (Control)
    Lysed Cells
    MCF7 Cells
    Lysate Type
    Cell Lysate
    Lysate Fraction
    Whole Cell Lysate
  • Indications d'application
    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.
    Commentaires

    Lysate Fractionation: Whole Cell Lysate
    Lysate Stimulation: Not Stimulated
    Lysate Tissue Culture: Tissue Culture

    Restrictions
    For Research Use only
  • Format
    Liquid
    Concentration
    1.0 mg/mL
    Buffer
    1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8)
    Conseil sur la manipulation
    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.
    Stock
    -80 °C
    Date de péremption
    3 months
  • Sujet
    Ready-to-use whole cell lysates produced 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.
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