A431 Cell Nuclear Extract
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- 宿主
- 人
- 资源
- 人细胞
- 应用范围
- Western Blotting (WB)
- 特异性
- 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.
- 产品特性
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Cell Line: Human A431 (epidermoid carcinoma)
Induction: None (Control) - 过滤
- Sterile filtered
- Lysed Cells
- A431 Cells
- Lysate Type
- Cell Lysate
- Lysate Fraction
- Nuclear Extract
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VNN1 293T Cell Transient Overexpression Lysate(Denatured)
VNN1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
CYP26A1 293T Cell Transient Overexpression Lysate(Denatured)CYP26A1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
APLP2 293T Cell Transient Overexpression Lysate(Denatured)APLP2 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
BIRC3 293T Cell Transient Overexpression Lysate(Denatured)BIRC3 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
BAG1 293T Cell Transient Overexpression Lysate(Denatured)BAG1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
ATF7 293T Cell Transient Overexpression Lysate(Denatured)ATF7 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
PF4 293T Cell Transient Overexpression Lysate(Denatured)PF4 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
ZD52F10 Overexpression Lysate (Denatured)DMKN WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
CLUHP3 Overexpression Lysate (Denatured)WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
PGM3 293T Cell Transient Overexpression Lysate(Denatured)PGM3 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
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- 应用备注
- 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.
- 说明
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Lysate Fractionation: Nuclear Extract
Lysate Stimulation: Not Stimulated
Lysate Tissue Culture: Tissue Culture - 限制
- 仅限研究用
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- 状态
- Liquid
- 浓度
- 1.0 mg/mL
- 缓冲液
- 1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8)
- 注意事项
- 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.
- 储存条件
- -80 °C
- 有效期
- 3 months
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- 背景
- 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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