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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 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.
- 产品特性
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Cell Line: HeLa - Human epidermoid carcinoma
Induction: None (Control) - Lysed Cells
- HeLa Cells
- Lysate Type
- Cell Lysate
- Lysate Fraction
- Nuclear Extract
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BIRC3 293T Cell Transient Overexpression Lysate(Denatured)
BIRC3 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
BAG1 293T Cell Transient Overexpression Lysate(Denatured)BAG1 WB Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
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
THP-1 (human acute monocytic leukemia) whole cell lysate (non-denatured)IP, WB Whole Cell Lysate Cell Lysate THP-1 Cells 适用: 人
Rat liver tissue lysate (non-denatured)IP, WB Whole Tissue Lysate Tissue Lysate 适用: 大鼠
Rat brain tissue lysate (non-denatured)IP, WB Whole Tissue Lysate Tissue Lysate 适用: 大鼠
ZSCAN21 HEK293 Cell Transient Overexpression Lysate(Non-Denatured)Zscan21 WB, IP Whole Cell Lysate Overexpression Lysate HEK 293T Cells 适用: 人
OLIG1 HEK293 Cell Transient Overexpression Lysate(Non-Denatured)OLIG1 WB, IP 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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