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DNA甲基转移酶活性/抑制极易检测试剂盒(比色法)
DNA甲基化是通过在胞嘧啶环的5-碳上的一个甲基的共价加成而发生的,结果是5-甲基胞嘧啶。 这些甲基投射到DNA的主要沟槽中并抑制转录。 在人类DNA中,5-甲基胞嘧啶大约存在于1.5%的基因组DNA中,主要在CpG位点。 在0.3到2 kb的DNA片段上有一簇被称为CpG岛的CpG位点,通常发现于基因的启动子区域或附近,那里是转录开始的地方。 在大部分基因组DNA中,大多数CpG位点被严重甲基化。 然而,CpG岛在种系组织和正常体细胞的启动子中仍未甲基化,允许基因表达。 当基因启动子区域的CpG岛甲基化时,基因的表达受到抑制。 这种抑制可以通过直接抑制特定转录因子的结合而引起,也可以通过招募甲基- cpg结合蛋白及其相关的抑制染色质重塑活性而间接引起。 除了对基因转录的影响,DNA甲基化还涉及基因组印记,这是指一个基因的亲代起源特异性表达,以及染色质结构域的形成。 在正常和病变细胞中,DNA甲基化在几个不同的水平上受到控制。 甲基的添加是由一个酶家族进行的,DNA甲基转移酶(DNMTs)。 在基因启动子附近的染色质结构也影响DNA甲基化和转录活性。 DNA甲基化模式的建立和维持需要三个DNMTs (DNMT1、DNMT3A和DNMT3B)。 另外两种酶(DNMT2和DNMT3L)也可能具有更专门化但相关的功能。 DNMT1似乎负责维持已建立的DNA甲基化模式,而DNMT3A和DNMT3B似乎介导了新的或从头DNA甲基化模式的建立。 研究发现DNMT3L是一个催化活性不高的DNA甲基转移酶调节因子,对DNMT3A和DNMT3B的功能至关重要。 病变细胞(如癌细胞)的不同之处在于,DNMT1并不单独负责维持异常的基因高甲基化,而DNMT1和DNMT3B可能在这一功能上相互合作。 局部染色质结构也有助于控制DNA甲基化。
本试剂盒包含测量DNMT活性或抑制的所有必要试剂。在这种检测中,通用的DNMT底物被稳定地涂在微孔板孔上。DNMT酶从AdoMet转移一个甲基到胞嘧啶,使DNA底物甲基化,甲基化的DNA可以用一个抗5-甲基胞嘧啶抗体识别。甲基化的DNA的比例或数量与酶的活性成正比,然后可以通过类似ELISA的反应,在微孔板分光光度计中以450纳米的波长读取吸光度来测量。DNMT酶的活性与测量的光密度强度成正比。
合作开发的DNA甲基转移酶活性/抑制极易检测试剂盒(比色法)。这个工具可在一个微孔板中定量DNMT,具有如下的优势和特性:
- 减少的步骤,使整个过程可在2小时10分钟内完成
- 灵敏度高,检测下限可低至0.2 ng 的纯化酶,灵敏度更优
- 便捷性好,所有组分预先制备,无须放射性或特殊仪器
- 改进的试剂盒组成,使检测具有更大的“信号窗口”,减少重复之间的误差
- 条状联孔式微孔板设计使分析灵活用于手工或高通量分析
- 简单,可靠,一致的分析条件
产品组分
组分内容 |
型号:A-P-3139-48(48次) |
型号:A-P-3139-96(96次) |
储存温度 |
|
WB |
10x Wash Buffer |
14 ml | 28 ml | 4°C |
DAB | DNMT Assay Buffer | 4 ml | 8 ml | 室温 |
SAM |
S-adenosylmethionine,50X* |
60 ul | 120 ul | –20°C |
DEC | DNMT Enzyme Control,50ug/ml* | 6 ul | 12 ul | –20°C |
CA |
Capature Antibody,1000X* |
5 ul | 10 ul | 4°C |
SI | Signal Indicator,1000X* | 6 ul | 12 ul | –20°C |
ES |
Enhancer Solution,2000X* |
6 ul | 12 ul | –20°C |
DS | Developing Solution | 5 ml | 10 ml | 4°C |
SS | Stop Solution | 5 ml | 10 ml | 室温 |
|
8-Well Assay Strips(With Frame) |
6 | 12 | 4°C |
Adhesive Covering Film | 1 | 1 | 室温 | |
操作手册 |
|
1 |
1 |
室温 |
文件资源
文件下载 | 文件内容 | 资源说明 |
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A-P-3139-DNA甲基转移酶活性/抑制极易检测试剂盒(比色法) | 操作手册 |
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注意事项
保存建议 | 推荐蓝冰或冰袋运输。当您收到产品后,按照说明书建议保存各组分。 |
警告 | 本品仅供科研使用,请勿用于临床与诊断。 |
FAQ
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