MCF-7 Cell Nuclear Extract
Quick Overview for MCF-7 Cell Nuclear Extract (ABIN964042)
Protein Species
Application
Lysed Cells
Lysate Type
Lysate Fraction
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Species of Lysate
- Human Cells
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Supplier Product No.
- w09-001-a85
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Supplier
- Rockland
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Specificity
- The cells were grown in EMEM supplemented with 10% FBS (Fetal Bovine Serum). Cells were washed in PBS and 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, 0.5% sodium deoxycholic acid and 0.1% SDS 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 sodium fluoride, sodium orthovanadate, sodium pyrophosphate and -glycerophosphate were also added. Cell debris was removed by centrifugation. 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.
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Characteristics
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Cell Line: MCF7 - Human Epithelial (Mammary)
Induction: None (Control) -
Sterility
- Sterile filtered
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NLRP3 293T Cell Transient Overexpression Lysate(Denatured)
NLRP3 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
TNFRSF13C 293T Cell Transient Overexpression Lysate(Denatured)TNFRSF13C WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
ZMYND19 293T Cell Transient Overexpression Lysate(Denatured)ZMYND19 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
MRGPRX2 293T Cell Transient Overexpression Lysate(Denatured)MRGPRX2 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
DDIT4L 293T Cell Transient Overexpression Lysate(Denatured)DDIT4L WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
NXNL1 293T Cell Transient Overexpression Lysate(Denatured)NXNL1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
LOC124220 293T Cell Transient Overexpression Lysate(Denatured)ZG16B WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
TMEM42 293T Cell Transient Overexpression Lysate(Denatured)TMEM42 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
SIRPD 293T Cell Transient Overexpression Lysate(Denatured)SIRPD WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
RASGRP4 293T Cell Transient Overexpression Lysate(Denatured)RASGRP4 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
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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.
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Comment
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Lysate Fractionation: Nuclear Extract
Lysate Stimulation: Not Stimulated
Lysate Tissue Culture: Tissue Culture -
Restrictions
- For Research Use only
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Format
- Liquid
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Concentration
- 1.0 mg/mL
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Buffer
- 1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8)
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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.
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Storage
- -80 °C
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Expiry Date
- 3 months
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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.
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