HeLa Whole Cell Lysate (TNFa Stimulated)

Détails pour le produit réf. ABIN1045084, Fournisseur: Connectez-vous pour afficher
Espèce de la protéine
Humain
Espèce du lysat
Human Cells
Application
Western Blotting (WB)
Options
Fournisseur
Connectez-vous pour afficher
N° du produit (Fournisseur)
Connectez-vous pour afficher
Specificité The cells were grown in Dulbecco’s medium supplemented with 10% fetal bovine serum. Cells were treated with 0.2 µg/ml TNF-alpha for 30 min. 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: HeLa - Human epidermoid carcinoma
Induction: Tumor Necrosis Factor-alpha (0.2 µg/ml)
Lysate Fraction Whole Cell Lysate
Lysate Treatment TNF-α stimulated
Lysate Type Cell Lysate
Lysed Cells HeLa Cells
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.
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: TNF alpha
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
Images (Fournisseur)
SDS-PAGE (SDS) image for HeLa Whole Cell Lysate (TNFa Stimulated) (ABIN1045084) Coommassie stained SDS-PAGE of 35 µg of TNF-a Stimulated HeLa Whole Cell Lysate (Read...