A431 Cell Nuclear Extract
Quick Overview for A431 Cell Nuclear Extract (ABIN964034)
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-a60
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Supplier
- Rockland
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Specificity
- The cells were grown in DMEM supplemented with 10% FBS (Fetal Bovine Serum). The lysate was prepared by first washing the cells in PBS. Washed cells were then incubated on ice in lysis buffer containing 10 mM HEPES, 60 mM KCl, 1.0 mM EDTA, 0.075% (v/v) NP40 and 1.0 mM DTT, pH 7.6. 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). Nuclei were then collected and washed in lysis buffer minus detergent. Nuclei were lysed by vortexing in extraction buffer containing 20 mM Tris-Cl, 1.5 mM MgCl2, 0.42 M NaCl, 0.2 mM EDTA, and 25% (v/v) glycerol, pH 8.0, supplemented with protease inhibitors (see above). The lysate was clarified by centrifugation. Protein concentration was determined by Lowry assay using a commercially available kit. The protein concentration was adjusted to 2.0 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: Human A431 (epidermoid carcinoma)
Induction: None (Control) -
Sterility
- Sterile filtered
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C1orf54 (1-131) 293T Cell Transient Overexpression Lysate(Denatured)
C1orf54 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
ID3 293T Cell Transient Overexpression Lysate(Denatured)ID3 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
HDAC1 293T Cell Transient Overexpression Lysate(Denatured)HDAC1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
HSPD1 293T Cell Transient Overexpression Lysate(Denatured)HSPD1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
HAL 293T Cell Transient Overexpression Lysate(Denatured)HAL WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
GYPA 293T Cell Transient Overexpression Lysate(Denatured)GYPA WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
MEIS1 293T Cell Transient Overexpression Lysate(Denatured)MEIS1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
IMPDH1 293T Cell Transient Overexpression Lysate(Denatured)IMPDH1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
HLA-E 293T Cell Transient Overexpression Lysate(Denatured)HLA-E WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells Reactivity: Human
MDM4 293T Cell Transient Overexpression Lysate(Denatured)MDM4 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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