检测多种物种中的m6A RNA去甲基化酶活性/抑制分析检测(哺乳动物,植物,真菌和细菌);2. 使用m6A抗体的比色ELISA样测定;3. 检出限低至2ug 核提取物和50ng纯化的酶。 3.m6A RNA甲基化抗体 m6A RNA甲基化抗体,#A-1801-020 图左:The N6-methyladenosine (m6A) Polyclonal Antibody tested in dot blot against ...
N6-methyladenosine(M6A)是在真核生物中最普遍的内部信使RNA(mRNA)修饰,并且在转录后调控中扮演关键角色。今年1月19日,南开大学药用化学生物学国家重点实验室和天津医科大学肿瘤研究所的研究人员在JBC Papers in Press上以“MicroRNA-145...
总RNA/mRNA m6A/ac4C定量检测(Dot Blot法) 在斑点杂交中,要检测到的生物分子不是首先通过电泳分离出来的,相反,将含有被检测分子的混合物直接点在膜上,烘烤固定。然后与m6A或Ac4C抗体(Western blot)杂交后检测,步骤如下: (1)交联:在65℃下5分钟,使mRNA变性以破坏RNA中的二级结构。变性后立即在冰上冷却,以防止...
m6A stabilizes RNA by 0.1–1.2 kcal mol−1most likely due to favorable stacking interactions between the methyl group and adjacent bases. Here, by attempting
首先利用MeRIP-seq测序数据结合m6A位点预测网站初步判断m6A修饰位点可能位于PxJHE的3′-UTR区靠近终止密码子附近,后续利用m6A-IP-qPCR技术确定了此m6A位点的真实性。利用双荧光素酶报告实验(Dual-luciferase reporter assays)和mRNA稳定性实验(mRNA stability assay)证明了此m6A位点负调控PxJHE基因表达。并且PxMettl3和Px...
RNA demethylases are α-ketoglutarate-dependent dioxygenases that can be inhibited by DMOG19. Such inhibition would be expected to increase the steady-state levels of m6A. We tested this possibility using dot blot assays on poly(A) RNA purified from Jurkat cells treated with 0.5 mM DMOG (...
1. 斑点杂交(Dot blot) 在研究 RNA 修饰时经常用到斑点杂交来对细胞内总RNA的修饰水平进行定性或半定量分析。具体步骤是指将待测的 RNA 变性后点加在硝酸纤维素膜或NC 膜上,紫外交联固定后用m6A或者m5C抗体进行杂交,洗膜,放射自显影,以斑点有无或颜色的深浅代表RNA修饰水平的高低。 (Sun B, et al. Plos ...
1. 斑点杂交(Dot blot) 在研究 RNA 修饰时经常用到斑点杂交来对细胞内总RNA的修饰水平进行定性或半定量分析。具体步骤是指将待测的 RNA 变性后点加在硝酸纤维素膜或 NC 膜上,紫外交联固定后用m6A或者m5C抗体进行杂交,洗膜,放射自显影,以斑点有无或颜色的深浅代表RNA修饰水平的高低。
Using mass spectrometry and dot blot assays, we demonstrated that m6A-RNA is actually present in the oyster and displays variations throughout early oyster development, with the lowest levels at the end of cleavage. In parallel, by in silico analyses, we were able to characterize at the ...
(D) Dot-blot assay of RNA m6A methylation in MDA-MB-231, MCF-7, and T47-D breast cancer cells overexpressing PRMT5 treated with doxorubicin as indicated. MDA-MB-231, 0.4 μg/mL DOX for 24 h; MCF-7 and T47-D, 0.5 μg/mL DOX for 24 h. (E) Dot-blot assay of RNA m6A ...