重组Anti-NRF1抗体[EPR5554(N)] - ChIP Grade (ab175932)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5554(N)] to NRF1 - ChIP Grade
- Suitable for: Flow Cyt (Intra), ChIP, WB, ICC/IF, IP, ChIC/CUT&RUN-seq, IHC-P, ChIP-sequencing
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-NRF1抗体[EPR5554(N)] - ChIP Grade
参阅全部 NRF1 一抗 -
描述
兔单克隆抗体[EPR5554(N)] to NRF1 - ChIP Grade -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ChIP, WB, ICC/IF, IP, ChIC/CUT&RUN-seq, IHC-P, ChIP-sequencingmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human NRF1 aa 350-450 (Cysteine residue). The exact sequence is proprietary.
Database link: Q16656 -
阳性对照
- WB: MCF-7, HeLa and 293T cell lysates and human fetal heart, mouse heart, mouse brain, rat heart and rat brain tissue lysates. IHC-P: Human gastric adenocarcinoma, human cervical carcinoma and human skeletal muscle tissues. ICC/IF: HeLa and MCF-7 cells. Flow Cyt (intra): 293T cells. IP: 293T cell lysate. ChIP-Seq: HeLA cells.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR5554(N) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab175932于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
1/10 - 1/150.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ChIP |
Use at an assay dependent concentration.
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WB | (2) |
1/1000 - 1/10000. Predicted molecular weight: 54 kDa.
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ICC/IF |
1/50 - 1/100.
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IP |
1/10 - 1/100.
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5 µg |
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ChIP-sequencing |
Use 8µg for 107 cells.
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说明 |
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Flow Cyt (Intra)
1/10 - 1/150. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ChIP
Use at an assay dependent concentration. |
WB
1/1000 - 1/10000. Predicted molecular weight: 54 kDa. |
ICC/IF
1/50 - 1/100. |
IP
1/10 - 1/100. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5 µg |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ChIP-sequencing
Use 8µg for 107 cells. |
靶标
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功能
Transcription factor that activates the expression of the EIF2S1 (EIF2-alpha) gene. Links the transcriptional modulation of key metabolic genes to cellular growth and development. Implicated in the control of nuclear genes required for respiration, heme biosynthesis, and mitochondrial DNA transcription and replication. -
组织特异性
Ubiquitously expressed with strongest expression in skeletal muscle. -
序列相似性
Belongs to the NRF1/Ewg family. -
翻译后修饰
Phosphorylation enhances DNA binding. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 4899 Human
- Entrez Gene: 18181 Mouse
- Entrez Gene: 312195 Rat
- Omim: 600879 Human
- SwissProt: Q16656 Human
- SwissProt: Q9WU00 Mouse
- SwissProt: Q62792 Rat
- Unigene: 654363 Human
see all -
别名
- alpha pal antibody
- alpha palindromic binding protein antibody
- Alpha palindromic-binding protein antibody
see all
图片
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 5 µg of ab175932 [EPR5554(N)]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The ChIP data was conducted on chromatin prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HeLa cells and 8 µg of ab175932. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HeLa cells and 8 µg of ab175932 [EPR5554(N)]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Additional screenshots of mapped reads can be downloaded here.
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All lanes : Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/5000 dilution (purified)
Lane 1 : Mouse heart tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Rat heart tissue lysate
Lane 4 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 54 kDa
Observed band size: 68 kDa why is the actual band size different from the predicted?
Blocking and dilution buffer: 5% NFDM/TBST -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cevical carcinoma tissue labelling NRF1 with purified ab175932 at a dilution of 1/100. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling NRF1 with purified ab175932 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
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Intracellular Flow Cytometry analysis of 293T cells labelling NRF1 with purified ab175932 at a dilution of 1/150 (red). Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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Chromatin was prepared from Hela cells according to the Abcam Dual X-ChIP protocol. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab175932 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
ab175932 (purified) at a dilution of 1/50 immunoprecipitating NRF1 in 293T whole cell lysate.
Lane 1 (input): 293T whole cell lysate (10µg)
Lane 2 (+): ab175932 + 293T whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab175932 in 293T whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Chromatin was prepared from NIH/3T3 treated with MG-132(2uM 16h) cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab175932 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/10000 dilution (purified) + HEK293 whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 68 kDa why is the actual band size different from the predicted?
Blocking and dilution buffer: 5% NFDM/TBST -
Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/10000 dilution (purified) + HeLa whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 68 kDa why is the actual band size different from the predicted?
Blocking and dilution buffer: 5% NFDM/TBST -
All lanes : Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/1000 dilution (unpurified)
Lane 1 : MCF-7 cell lysate
Lane 2 : Hela cell lysate
Lane 3 : Human fetal heart tissue lysate
Lane 4 : 293T cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 54 kDa -
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labeling NRF1 with unpurified ab175932 at a dilution of 1/50.
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ab175932 (unpurified) at a dilution of 1/10 immunoprecipitating NRF1 in 293T cell lysate.
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Intracellular flow cytometric analysis of permeabilized 293T cells labeling NRF1 with unpurified ab175932 at a dilution of 1/10 (red) compared to a negative control (rabbit IgG,green).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue labeling NRF1 with unpurified ab175932 at a dilution of 1/50.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric adenocarcinoma tissue labeling NRF1 with unpurified ab175932 at a dilution of 1/50.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (57)
ab175932 被引用在 57 文献中.
- He K et al. Hippocampus-Based Mitochondrial Respiratory Function Decline Is Responsible for Perioperative Neurocognitive Disorders. Front Aging Neurosci 14:772066 (2022). PubMed: 35221986
- Zhang LQ et al. 5-HT1F Receptor Agonist Ameliorates Mechanical Allodynia in Neuropathic Pain via Induction of Mitochondrial Biogenesis and Suppression of Neuroinflammation. Front Pharmacol 13:834570 (2022). PubMed: 35308244
- Niu N et al. Effects of NRF-1 and PGC-1α cooperation on HIF-1α and rat cardiomyocyte apoptosis under hypoxia. Gene 834:146565 (2022). PubMed: 35569770
- Liu X et al. LINC00839 promotes colorectal cancer progression by recruiting RUVBL1/Tip60 complexes to activate NRF1. EMBO Rep 23:e54128 (2022). PubMed: 35876654
- Zhang L et al. LncRNA NR2F1-AS1 Inhibits the Malignant Properties of Cervical Cancer Cells via Targeting miR-642a-3p/NR2F1 Axis. Rev Invest Clin 74:181-192 (2022). PubMed: 36087939