| 货号 | 12721S |
| 反应种属 | Human/Mouse/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/F/ChIP |
| 使用方法 | WB(1:1000) IP (1:50) F (1:3200) ChIP (1:100) |
| 供应商 | CST |
| 背景 | The nuclear factor-like 2 (NRF2) transcriptional activator binds antioxidant response elements (ARE) of target gene promoter regions to regulate expression of oxidative stress response genes. Under basal conditions, the NRF2 inhibitor INrf2 (also called KEAP1) binds and retains NRF2 in the cytoplasm where it can be targeted for ubiquitin-mediated degradation (1). Small amounts of constitutive nuclear NRF2 maintain cellular homeostasis through regulation of basal expression of antioxidant response genes. Following oxidative or electrophilic stress, KEAP1 releases NRF2, thereby allowing the activator to translocate to the nucleus and bind to ARE-containing genes (2). The coordinated action of NRF2 and other transcription factors mediates the response to oxidative stress (3). Altered expression of NRF2 is associated with chronic obstructive pulmonary disease (COPD) (4). NRF2 activity in lung cancer cell lines directly correlates with cell proliferation rates, and inhibition of NRF2 expression by siRNA enhances anti-cancer drug-induced apoptosis (5). |
| 存放说明 | -20C |
| 计算分子量 | 97-100 |
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。 |
