Cell Signaling Technology

Product Pathways - Transcription Factors

NRF2 (D1Z9C) XP® Rabbit mAb #12721

sc-13032   sc-722  

No. Size Price
12721S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
12721T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
12721 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Monkey, Endogenous 97-100 Rabbit IgG
IP 1:50
F 1:3200
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, ChIP=Chromatin IP,

Specificity / Sensitivity

NRF2 (D1Z9C) XP® Rabbit mAb recognizes endogenous levels of total NRF2 protein.

NRF2 (D1Z9C) XP® Rabbit mAb 能够检测内源性NRF2总蛋白水平。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala275 of human NRF2 protein.

该单克隆抗体是经由合成的围绕人NRF2蛋白Ala275位点周围相应的多肽片段免疫动物而生产的。

Chromatin IP

Chromatin IP

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 Cytometry

Flow Cytometry

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 Blotting

Western Blotting

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。

IP

IP

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。

Background

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).

Nuclear factor-like 2 (NRF2)转录激活因子能够结合靶基因启动子区域的antioxidant response elements (ARE),从而调节氧化应激反应基因的表达。在生理条件下,NRF2抑制因子INrf2 (又称KEAP1)结合NRF2并使NRF2保持在细胞质中,从而使NRF2通过泛素化通路介导降解(1)。少量组成型NRF2通过抗氧化反应基因的基本表达的调控来维持细胞内稳定。在氧化应激或亲电子反应后,KEAP1蛋白释放NRF2蛋白,从而允许该转录激活因子进入细胞核,并且结合到靶基因的ARE元件(2)。NRF2蛋白和其它转录因子协同介导氧化应激反应(3)。NRF2蛋白的可变表达与chronic obstructive pulmonary disease (COPD)有关(4)。在肺癌细胞系中NRF2蛋白的活性直接与细胞增殖率有关,同时通过siRNA沉默抑制NRF2蛋白表达能够提高抗癌药物介导的凋亡(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.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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