HeLa Whole Cell Lysate

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Proteinspezies
Human
Lysatspezies
Human Cells
Applikation
Western Blotting (WB)
Optionen
Hersteller
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Spezifität The Hela 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.
Produktmerkmale Cell Line: HeLa - Human epidermoid carcinoma
Induction: None (Control)
Lysate Fraction Whole Cell Lysate
Lysate Type Cell Lysate
Lysed Cells HeLa Cells
Hintergrund Ready-to-use HeLa 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.
Synonyms: Hela Cells, HeLa Lysate, Whole Cell Lysate
Applikations-hinweise Ready-to-use HeLa whole cell 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.
Kommentare

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

Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 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)
Handhabung Ready-to-use HeLa whole cell 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.
Lagerung -80 °C
Haltbarkeit 3 months
Bilder des Herstellers
 image for HeLa Whole Cell Lysate (ABIN964024) HeLa Whole Cell Lysate
Western Blotting (WB) image for HeLa Whole Cell Lysate (ABIN964024) Western Blot showing detection of alpha tubulin in lane 1. HeLa Whole Cell Lysate (10...
SDS-PAGE (SDS) image for HeLa Whole Cell Lysate (ABIN964024) Coommassie stained SDS-PAGE of 10 µg of Human Derived HeLa Whole Cell Lysate separate...