Protein Species: Human
Species of Lysate: Human Cells
Application: Western Blotting (WB)
Specificity: 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.
Characteristics: Cell Line: HeLa - Human epidermoid carcinoma
Induction: None (Control)
Lysed Cells: HeLa Cells
Lysate Type: Cell Lysate
Lysate Fraction: Nuclear Extract

BIRC3 293T Cell Transient Overexpression Lysate(Denatured)

BIRC3 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human

APLP2 293T Cell Transient Overexpression Lysate(Denatured)

APLP2 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human

BAG1 293T Cell Transient Overexpression Lysate(Denatured)

BAG1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human

VNN1 293T Cell Transient Overexpression Lysate(Denatured)

VNN1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human

CYP26A1 293T Cell Transient Overexpression Lysate(Denatured)

CYP26A1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human

Hep2 Whole Cell Lysate

WB Whole Cell Lysate Cell Lysate HepG2 Cells Reactivity: Human

NIH/3T3 Whole Cell Lysate (PDGF Stimulated)

WB Whole Cell Lysate Cell Lysate 3T3 Cells PDGF Stimulated Reactivity: Mouse

DYKDDDDK Positive Control Lysate

PC, WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Tag

A431 Whole Cell Lysate (EGF Stimulated)

WB Whole Cell Lysate Cell Lysate A431 Cells EGF treated Reactivity: Human

CD177 (human): 293T Lysate

CD177 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells

Application Notes: Ready-to-use nuclear extracts are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Nuclear extracts are supplied in denaturing buffer without dissociating agents. Heat nuclear extract 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 10-30 l depending on the size format of your gel.
Comment: Lysate Fractionation: Nuclear Extract
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)
Handling Advice: Ready-to-use nuclear extracts are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Nuclear extracts are supplied in denaturing buffer without dissociating agents. Heat nuclear extract 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 10-30 µl depending on the size format of your gel.
Storage: -80 °C
Expiry Date: 3 months

Background: Ready-to-use nuclear extracts 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.

HeLa Cell Nuclear Extract

Product Details