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Souris Embryonic Fibroblast Whole Cell Lysate

WB Whole Cell Lysate Cell Lysate Fibroblast Cells Reactivité: Souris
Rockland
N° du produit ABIN964051
N° du produit (Fournisseur): w10-001-371

Aperçu rapide pour Souris Embryonic Fibroblast Whole Cell Lysate (ABIN964051)

Espèce de la protéine

Souris

Application

Western Blotting (WB)

Lysed Cells

Fibroblast Cells

Lysate Type

Cell Lysate

Lysate Fraction

Whole Cell Lysate
  • Espèce du lysat

    Souris

    N° du produit (Fournisseur)

    w10-001-371

    Fournisseur

    Rockland

    Specificité

    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 containing 150 mM sodium chloride, 50 mM Tris Cl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.25% sodium deoxycholic acid 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 2X SDS-PAGE sample buffer was added.

    Attributs du produit

    Cell Line: Mouse Embryonic Fibroblast
    Induction: None (Control)

    Stérilité

    Sterile filtered
  • 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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