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Product Usage Information
For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Immunofluorescence (Immunocytochemistry) | 1:200 - 1:800 |
| Flow Cytometry (Fixed/Permeabilized) | 1:1600 - 1:6400 |
| Chromatin IP | 1:200 |
| Chromatin IP-seq | 1:200 |
Specificity/Sensitivity
物种反应性：
人, 小鼠, 猴
核因子样 2 (NRF2) 转录激活因子结合靶标基因启动子区域的抗氧化反应元件 (ARE)，从而调控氧化应激反应基因的表达。在基础条件下，NRF2 抑制剂 INrf2（也称为 KEAP1）结合 NRF2，并将其保留在细胞浆内，以便在细胞浆内靶向进行泛素介导的降解 (1)。少量组成型胞核 NRF2 通过调节抗氧化剂反应基因的基础表达来维持细胞稳态。在氧化或亲电子应激后，KEAP1 释放 NRF2，从而使激活剂转位到细胞核与 ARE 元件基因结合 (2)。NRF2 与其他转录因子的协同作用可介导氧化应激反应 (3)。NRF2 的表达改变与慢性阻塞性肺疾病 (COPD) 相关 (4)。肺癌细胞系的 NRF2 活性与细胞增殖率直接相关，并且 siRNA 对 NRF2 表达的抑制会增强抗癌药物的诱导性凋亡 (5)。 1.Cullinan, S.B. et al. (2004) Mol Cell Biol 24, 8477-86. 2.Nguyen, T. et al. (2005) J Biol Chem 280, 32485-92. 3.Jaiswal, A.K. (2004) Free Radic Biol Med 36, 1199-207. 4.Suzuki, M. et al. (2008) Am J Respir Cell Mol Biol 39, 673-82. 5.Homma, S. et al. (2009) Clin Cancer Res 15, 3423-32.
参考图片
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MEF NRF2 wild-type (left) and NRF2 knock-out (right) cells, both treated with DEM (50 μM, 3 hr), and 5 µl of NRF2 (D1Z9C) XP® Rabbit mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using mouse MafG intron 1 primers, SimpleChIP® Mouse NQO1 Promoter Primers #12635, and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
使用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 分别对4 x 106 MEF NRF2 野生型 （左）以及NRF2 敲除（右）细胞中交联过的染色质进行染色质免疫沉淀，所用抗体为5 µl NRF2 (D1Z9C) XP® Rabbit mAb兔单抗 或2 µl Normal Rabbit IgG #2729。富集到的DNA经过实时PCR定量，所使用产品为MafG intron 1 primers， SimpleChIP® Mouse NQO1 Promoter Primers #12635以及 SimpleChIP® Mouse RPL30 Intron 2 Primers #7015。每个样品中免疫沉淀得到的DNA量由相对于input染色质总量( 相当于1)的信号值来表示。
Flow cytometric analysis of MEF wt cells, untreated (blue) and treated with MG-132 #2194 (green), using NRF2 (D1Z9C) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.
流式细胞术分别检测未处理（蓝色）或经MG-132 #2194处理（绿色）的MEF wt细胞活性，使用的抗体为NRF2 (D1Z9C) XP® Rabbit mAb兔单抗，所用二抗为F(ab')2Fragment (Alexa Fluor® 647接合的) #4414。
Western blot analysis of extracts from MEF wt and U-2 OS cells, untreated (-) or treated with MG-132 #2194 (10 μM, 10 hr; +), using NRF2 (D1Z9C) XP® Rabbit mAb.
Western blot方法分别检测未处理（-）或经MG-132 #2194 (10 μM, 10 hr; +)处理的MEF wt和 U-2 OS细胞提取物，使用的抗体为NRF2 (D1Z9C) XP® Rabbit mAb兔单抗。
Immunoprecipitation of NRF2 from MEF wt cell extracts treated with MG-132 #2194 (10 μM, 10 hr) using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or NRF2 (D1Z9C) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using NRF2 (D1Z9C) XP® Rabbit mAb (lane 3).
经MG-132 #2194 (10 μM, 10 hr)处理的MEF wt细胞提取物中免疫沉淀NRF2，使用的抗体为Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) 或NRF2 (D1Z9C) XP® Rabbit mAb兔单抗 (lane 3)。Lane 1为10% input。 使用NRF2 (D1Z9C) XP® Rabbit mAb兔单抗 (lane 3)进行Western blot检测。
危险品化学品经营许可证(不带存储) 许可证编号：沪(杨)应急管危经许[2022]202944(QY) 
营业执照(三证合一)