細胞核/細胞質分離試劑盒 | Nuclear/Cytosol Fractionation Kit貨號K266-100 (ABCAM, ab289882)

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特點

胞器蛋白質分離–系列商品介紹

細胞核/細胞質分離試劑盒 Nuclear/Cytosol Fractionation Kit | 貨號K266-100(ABCAM, ab289882)

Nuclear/Cytosol Extraction Kit試劑盒內包含，分離哺乳動物細胞的細胞核和細胞質所有試劑，不僅能夠分離細胞核和細胞質蛋白質，而且可以保持蛋白質的完整性。所分離的細胞核和細胞質蛋白質可進行轉錄活性(transcriptional activity)、RNA 剪接 (RNA splicing)、凝膠阻滯電泳實驗 (gel shift assay)、報告蛋白檢測 ( reporter assays)、酶活性檢測 (enzyme activity assays)及Western Blot 研究等。

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Detection Method	N/A
Species Reactivity	Mammalian
Applications	This Nuclear/Cytosol Extraction Kit provides a complete system that enables the separation of nuclear extract from the cytoplasmic fraction of mammalian cells and tissues The new kit provides a system that maintains the nuclear and cytoplasmic compartments separate and intact. Transcriptional activity/reporter assays/enzyme activity assays/Western blotting and more downstream assays.
Features & Benefits	• Simple procedure; takes less than 2-3 hours • Fast and convenient • The optimized reagents and procedures provided with the kit allow separation of nuclear and cytoplasmic extracts from cells relatively quick with little or no cross-contaminations
Kit Components	• Cytosol Extraction Buffer A (CEB-A) • Cytosol Extraction Buffer B (CEB-B) • Nuclear Extraction Buffer (NEB) • DTT (1 M) • Protease Inhibitor Cocktail (lyophilized)

胞器蛋白分離的應用

探討細胞核內轉錄因子和特定DNA序列相互作用。檢測蛋白質和一段特定DNA序列(label)相互結合的技術原理: 當特定序列的DNA和某蛋白質結合後，整個蛋白質複合體分子量會增加,經由電泳後,在gel中的移動速度變慢,整個band進上可加入特定抗體檢測是否有特定蛋白質Binding( Supershift)
細胞凋亡時,粒線體膜電位去極化的過程會增加膜的通透性並使膜內蛋白,如Cytochrome C, AIF, Smac/DIABLO等分子釋放到細胞質中,引發caspase-9活化, 造成一連串凋亡反應發生. COX4蛋白位於粒腺體細胞內側, 並在Apoptosis發生時依舊保留在粒線體內。
二維電泳與MDLC(液相層析)並列為研究蛋白質體學的重要技術之一。原理: 利用分子量(一維)與等電點(二維)一次可分離大約2000個蛋白質。限制: 膜蛋白(Hydrophobic)在電泳中不易看到.蛋白質若是為low abundant protein則欲得到準確而可信的實驗結果要做胞器分離( sample prefractionation) 胞器分離在2d應用的優點: 移去abundant protein 、增加 low abundant protein、減少蛋白質的複合體發生

引用文獻

Stephen A. Martin, et al., (2018) GSK3β Regulates Brain Energy Metabolism, Cell Reports, May 2018, 29768193

Liu, Xiang-Qian et al. (2017) Inhibitory effects of 3α-hydroxy-lup-20(29)-en-23, 28-dioic acid on lipopolysaccharide-induced TNF-α, IL-1β, and the high mobility group box 1 release in macrophages, Biosci Biotechnol Biochem. 2017 Jul;81(7):1305-1313.

Chen, Xiuqing et al. (2017) Attenuation of acute lung injury in a rat model by Semen Cassiae, BMC Complement Altern Med. 2017 Apr 28;17(1):234.

Cho, Haaglim et al. (2017) ENOblock, a unique small molecule inhibitor of the non-glycolytic functions of enolase, alleviates the symptoms of type 2 diabetes, Sci Rep. 2017 Mar 8;7:44186.

Oh, Sea-Hun et al. (2017) 4-hydroxybenzaldehyde-chitooligomers suppresses H2O2-induced oxidative damage in microglia BV-2 cells, Carbohydr Res. 2017 Feb 22;440-441:32-37.

產品編號	產品名稱	包裝
K270-50	FractionPREP™ Cell Fractionation Kit 四種胞器萃取一組搞定	50 assays
K256-25 K256-100	Mitochondria/Cytosol Fractionation Kit	25 assays 100 assays
K266-25 K266-100	Nuclear/Cytosol Fractionation Kit	25 assays 100 assays
K267-50	Cytosol/Particulate Separation Kit	50 assays
K269-500	Mammalian Cell Extraction Kit	500 assays
K268-50	Membrane Protein Extraction Kit	50 assays
K288-50	Mitochondria Isolation Kit For Tissue & Cultured Cells	50 assays
K280-50	Mitochondrial DNA Isolation Kit	50 assays
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