核Extraction试剂盒- Soluble / Insoluble Fractions (ab219177)
Key features and details
- Assay time: 45 min
- Sample type: Adherent cells, Suspension cells, Tissue
概述
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产品名称
核Extraction试剂盒- Soluble / Insoluble Fractions
参阅全部 Nuclear Extraction 试剂盒 -
样品类型
Tissue, Adherent cells, Suspension cells -
检测时间
0h 45m -
产品概述
Nuclear extraction kit ab219177 offers a simple 45 minute method for extracting nuclear proteins from mammalian cells or tissues.
For optimal extraction of nuclear proteins, the kit produces three distinct extracts (fractions): soluble nuclear proteins (nuclear extract 1), nuclear proteins that remain insoluble after production of nuclear extract 1 (nuclear extract 2), and cytoplasmic proteins.
The kit contains reagents for 100 extractions, where each extraction starts with 5 x 106 cells or 50 mg of tissue. Typical yields from each extraction are 0.3 – 0.4 mg of soluble nuclear proteins, 0.1 – 0.2 mg of insoluble nuclear proteins and 0.6 – 0.7 mg of cytoplasmic proteins. Nuclear proteins extracted with this kit can be used in a variety of applications such as western blotting and nuclear enzyme assays.
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说明
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
性能
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存放说明
Store at +4°C. Please refer to protocols. -
组件 100 tests 200X DTT 1 x 1ml Cytoplasm Extraction Buffer 1 x 55ml Nuclear Extraction Buffer 1 x 55ml Nuclear Lysis Buffer 1 x 55ml Protease Inhibitor Cocktail 1 x 1ml
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图片
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Extracts were prepared from 5 x 106 HeLa (human epithelial cell line from cervix adenocarcinoma) cells using Nuclear Extract Kit ab219177 and analysed by SDS-PAGE with Coomasie Blue staining. Extract or whole cell lysate from 3 x 104 cells was loaded in each lane along with BSA standards. Lanes: 1. MW marker; 2. Whole cell lysate; 3. Cytoplasmic extract; 4. Nuclear extract 1; 5. Nuclear extract 2; 6. MW marker; 7. BSA, 2.0 µg; 8. BSA 1.0 µg; 9. BSA 0.5 µg; 10. BSA 0.2 µg; 11. BSA 0.1 µg.
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Extracts were prepared from HeLa (human epithelial cell line from cervix adenocarcinoma) cells using the manufacturer’s protocol and analyzed by Western blot. For each manufacturer: (1) cytoplasmic extract, (2) nuclear extract, (3) nuclear extract 2 or resolubilized insoluble nuclear fraction.
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Extracts were prepared from 5 x 106 HeLa (human epithelial cell line from cervix adenocarcinoma) cells using Nuclear Extract Kit ab219177. Note that protease inhibitors were omitted from all buffers to prevent interference with the HDAC activity assay. The enzymatic activity of HDACs in each fraction was measured using the Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) (ab156064). Extract from of 3 x 104 cells was used per assay test.
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Extracts were prepared from 5 x 106 cells from RAJI (human lymphoblastoid cell line), HEK-293 (human embryonic kidney cell line), SHSY-5Y (human neuroblastoma cell line) and U87-MG (human glioblastoma cell line) cell lines using Nuclear Extract Kit ab219177 and analysed by Western blot. Extract from 3 x 104 cells was loaded per lane.
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Extracts were prepared from 120 mg of mouse tissue using Nuclear Extract Kit ab219177 and analysed by Western blot. Extract from 0.4 mg of tissue was loaded per lane.
数据表及文件
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SDS download
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Datasheet download
文献 (4)
ab219177 被引用在 4 文献中.
- Bielczyk-Maczynska E et al. G protein-coupled receptor 151 regulates glucose metabolism and hepatic gluconeogenesis. Nat Commun 13:7408 (2022). PubMed: 36456565
- Turnbull RE et al. The MiDAC histone deacetylase complex is essential for embryonic development and has a unique multivalent structure. Nat Commun 11:3252 (2020). PubMed: 32591534
- Bautista SJ et al. mTOR complex 1 controls the nuclear localization and function of glycogen synthase kinase 3ß. J Biol Chem 293:14723-14739 (2018). PubMed: 30061153
- Pascual-Reguant L et al. Lamin B1 mapping reveals the existence of dynamic and functional euchromatin lamin B1 domains. Nat Commun 9:3420 (2018). PubMed: 30143639