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: MCF7 - Human Epithelial (Mammary)
Induction: None (Control)
Lysed Cells: MCF7 Cells
Lysate Type: Cell Lysate
Lysate Fraction: Whole Cell Lysate

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

APLP2 293T Cell Transient Overexpression Lysate(Denatured)

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

BIRC3 293T Cell Transient Overexpression Lysate(Denatured)

BIRC3 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

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 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.
Comment: 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)
Handling Advice: 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.
Storage: -80 °C
Expiry Date: 3 months

Background: 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.

MCF-7 Whole Cell Lysate

Product Details