Mouse Embryonic Fibroblast Whole Cell Lysate
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- Protein Species
- Mouse
- Species of Lysate
- Mouse
- Application
- Western Blotting (WB)
- Specificity
- The cells were grown in RPMI supplemented with 10% FBS (Fetal Bovine Serum). The lysate was prepared by first washing the cells in PBS. Washed cells are then incubated on ice in modified RIPA buffer containing 150 mM sodium chloride, 50 mM Tris Cl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.25% sodium deoxycholic acid 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). Phosphatase inhibitors include 1 mM Sodium Fluoride and 1mM Na3VO4. Cell debris was removed by 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.
- Characteristics
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Cell Line: Mouse Embryonic Fibroblast
Induction: None (Control) - Sterility
- Sterile filtered
- Lysed Cells
- Fibroblast Cells
- Lysate Type
- Cell Lysate
- Lysate Fraction
- Whole Cell Lysate
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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 LysateWB 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 LysatePC, 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 LysateCD177 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells
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- 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
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Lysate Fractionation: Whole Cell Lysate
Lysate Stimulation: Not Stimulated
Lysate Tissue Culture: Tissue Culture - Restrictions
- For Research Use only
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- 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
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- 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.
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