5-羥甲基胞嘧啶(5-hydroxymethylcytosine,5-HT)作為第六種DNA堿基,在人類和小鼠的腦細胞和胚胎干細胞中含量豐富,具有轉錄調控功能。在哺乳動物中,它可以通過5-甲基胞嘧啶的氧化產生,這是由胞嘧啶加氧酶的10 - 11易位(TET)家族介導的反應。
胞嘧啶加氧酶的TET家族包括TET 1、TET 2和TET 3。這些泰特蛋白可以通過結合到富含CpG的區域以防止不需要的DNA甲基轉移酶活性,以及通過將5 mC轉化為5 hmC并通過羥化酶活性進一步轉化為5caC(5-羧基胞嘧啶)來促進DNA去甲基化。結果表明,基因組5 hmC水平與泰特羥化酶活性相關。此外,TET 1通過結合ES細胞中的不同靶基因而顯示出在轉錄激活和抑制中具有雙重功能。TET 1也是急性髓性白血病中MLL基因的融合伴侶,被認為是一種癌蛋白。發現TET 2在白血病中經常突變,并被認為是腫瘤抑制劑。TET 3被證明在哺乳動物受精卵中的DNA甲基化重編程過程中發揮獨特的作用。因此,激活腫瘤抑制泰特酶如TET 2或抑制癌蛋白TET酶如TET 1將在有益于癌癥診斷和開發新的基于靶點的癌癥治療劑方面是重要的。
那么如何檢測5hmc羥化酶TET的活性呢?
作為專業的生命科學解決方案供應商,艾美捷為您推薦Nature同款方案:
產品名稱 | 5hmC-羥化酶TET活性/抑制分析試劑盒 5mC-Hydroxylase TET Activity/Inhibition Assay Kit |
產品貨號 | P-3086,比色法 P-9087,熒光法 |
產品原理 | 在本實驗中,甲基化底物被穩定地包埋在微孔板上?;钚?TETs (客戶提供)與底物結合,并將甲基化底物轉化為羥甲基化產物。Tet- 轉化的羥甲基化產物可以被特異性抗體識別。羥甲基化產物的比例或數量與酶的活性成正比,然后可以通過讀取波長為450nm的吸光度或者熒光強度來對比分析。TET 酶的活性與測得的光強度成正比。 |
適用樣本 | TET酶的細胞核提取物;純化的TET酶(TETs, 1-3) |
性能展示 | |
說明書下載 | 點擊下載 |
5hmC-羥化酶TET活性/抑制分析試劑盒 優點和特點:
1、5小時比色測定程序,步驟簡單,方便快捷。整個過程可在5小時內完成。
2、通過直接檢測TET轉化的羥甲基化產物直接測量泰特羥化酶活性。
3、創新的試劑盒組成使背景信號極低,并允許檢測簡單、準確、可靠和一致。
4、可以使用細胞/組織提取物或純化泰特蛋白,其允許檢測TET羥化酶抑制劑在體內和體外的抑制作用。
5、新的測定原理允許實現高靈敏度?;钚钥梢詮牡椭?0ng的純化的TET1羥化酶檢測。
6、包括羥甲基化標準品,其允許定量泰特羥化酶的比活性。
7、條帶微孔板形式使得該測定方法靈活,可用于手動或高通量分析(96次測定)。
Nature文章結果:
DOI https://doi.org/10.1038/s41590-020-0764-8
1.TET酶活性分析使用P-3086; 使用艾美捷推薦的P-1032檢測整體5hmc水平
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