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Normal Poulet Spinal Cord Whole Cell Lysate

WB Whole Tissue Lysate Tissue Lysate Reactivité: Poulet
Rockland
N° du produit ABIN964018
N° du produit (Fournisseur): w03-001-t296
277,44 €
Frais d'envoi 40,00 €, glace carbonique 20,00 € et TVA exclus
200 μg
Destination: France
Envoi sous 5 à 7 jours ouvrables

Aperçu rapide pour Normal Poulet Spinal Cord Whole Cell Lysate (ABIN964018)

Espèce de la protéine

Poulet

Application

Western Blotting (WB)

Lysate Type

Tissue Lysate

Lysate Fraction

Whole Tissue Lysate
  • Espèce du lysat

    Poulet

    N° du produit (Fournisseur)

    w03-001-t296

    Fournisseur

    Rockland

    Specificité

    Tissues were washed exhaustively with PBS to remove blood and other debris. A lysate was prepared by homogenizing the tissue and washing the cells in cold PBS. Washed cells were incubated at 4° C in modified RIPA buffer containing 150 mM sodium chloride, 50 mM Tris Cl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.5% sodium deoxycholic acid and 0.1% SDS 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). The following phosphatase inhibitors were also added: 1 mM NaF and 1 mM Na3VO4. Cell debris was removed by centrifugation and 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: Primary tissue
    Induction: None (Control)
  • 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 5-15 µl depending on the size format of your gel.

    Commentaires

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

    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 5-15 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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