Name: Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade
SKU: ab175932
Rating: 4 (2 reviews)

Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) is a rabbit monoclonal antibody that is used to detect NRF1 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF, ChIP. Suitable for Human, Mouse, Rat samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

-Over 70 publications

Images

ChIP - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (AB175932), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ChIC/CUT&RUN-seq	IHC-P	IP	ChIP	WB	ICC/IF	ChIP-seq	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Expected	Expected	Tested	Tested	Expected	Expected	Expected
Rat	Expected	Expected	Expected	Expected	Tested	Expected	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 5 µg	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info 5 µg	Notes -
Species Rat	Dilution info 5 µg	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50 - 1/100	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse, Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/10000	Notes -
Species Rat	Dilution info 1/1000 - 1/10000	Notes -
Species Human	Dilution info 1/1000 - 1/10000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50 - 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 8 µg for 10⁷ Cells	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/150	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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3 products for Alternative Product

Target data

See full target information NRF1

Function

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.

Alternative names

Nuclear respiratory factor 1, NRF-1, Alpha palindromic-binding protein, Alpha-pal, NRF1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR5554(N)
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), ChIP in Human, Mouse, Rat samples.

What is the molecular weight of NRF1?

Anti-NRF1 [EPR5554(N)] - ChIP Grade (ab175932) specifically detects a band for NRF1 (UniProt: Q16656) at a molecular weight of 54kDa.

Trusted by the scientific community

Anti-NRF1 [EPR5554(N)] - ChIP Grade (ab175932) was first used in a scientific publication in 2013 and has been cited over 70 times in peer-reviewed journals.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products

We have a range of other formats of antibody clone [EPR5554(N)] also available for your convenience:
ab175932, Alexa Fluor® 488 - ab200976, Alexa Fluor® 647 - ab200979, Carrier free - ab221792

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The nuclear respiratory factor 1 (NRF1) also known as Alpha-Pal is a transcription factor with an approximate molecular weight of 68 kDa. It plays an important role in the regulation of genes required for mitochondrial DNA transcription and replication. Researchers find NRF1 expression in various tissues including muscle brain and liver. This protein functions mechanistically by binding to specific promoter regions of target genes to control their expression making it essential for maintaining cellular energy metabolism.

Biological function summary

NRF1 influences the expression of genes involved in the mitochondrial function and is part of the transcriptional complex. This complex includes the NRF2 protein where NRF1 partners with co-activators to modulate gene transcription. NRF1 promotes the expression of components essential for mitochondrial biogenesis and cellular respiration which are vital processes for energy production in cells. Therefore NRF1 contributes to maintaining cellular energy homeostasis and overall cellular health.

Pathways

NRF1 integrates into critical regulatory pathways for mitochondrial function and metabolism. NRF1 links closely to the PGC-1α pathway which is instrumental in the regulation of energy metabolism and mitochondrial biogenesis. Additionally NRF1 works in conjunction with NRF2 to regulate oxidative stress responses. These interactions ensure a coordinated cellular response to energy demands and help protect the cell from oxidative damage.

Associated diseases and disorders

NRF1 associates with neurodegenerative diseases and metabolic syndromes. Altered NRF1 regulation can impact diseases such as Alzheimer's disease where it may contribute to neurodegeneration through impaired mitochondrial function. Moreover the dysfunction of NRF1 might relate to metabolic disorders like diabetes as energy metabolism and mitochondrial health are disrupted. NRF1 connects with proteins like SIRT1 in these contexts further implicating NRF1 in these pathological states through its role in energy homeostasis and stress response.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

18 product images

ChIP - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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
ChIC/CUT&RUN sequencing - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ChIC/CUT&RUN pAG-MNase ab285373, 2.5x10^5 HeLa cells and 5μg of ab175932 [EPR5554(N)]. The resulting DNA was sequenced on the NovaSeq 6000 (PE150).
Additional screenshots of mapped reads can be downloaded here.
ChIP - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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
ChIP-sequencing - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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.
Immunoprecipitation - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab175932 in 293T whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (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.
All lanes: Immunoprecipitation - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
Predicted band size: 54 kDa
Observed band size: 68 kDa
Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
Blocking and dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/5000 dilution
Lane 1: Mouse heart tissue lysate at 20 µg
Lane 2: Mouse brain tissue lysate at 20 µg
Lane 3: Rat heart tissue lysate at 20 µg
Lane 4: Rat brain tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 54 kDa
Observed band size: 68 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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. Goat Anti-Rabbit IgG H&L (HRP) 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.
Immunocytochemistry/ Immunofluorescence - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).
Flow Cytometry (Intracellular) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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.
Immunoprecipitation - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
ab175932 (unpurified) at a dilution of 1/10 immunoprecipitating NRF1 in 293T cell lysate.
All lanes: Immunoprecipitation - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
Predicted band size: 54 kDa
Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
Blocking and dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/10000 dilution
All lanes: HEK293 whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 68 kDa
Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
Blocking and dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/10000 dilution
All lanes: HeLa whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 68 kDa
Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
All lanes: Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/1000 dilution
Lane 1: MCF-7 cell lysate at 10 µg
Lane 2: Hela cell lysate at 10 µg
Lane 3: Human fetal heart tissue lysate at 10 µg
Lane 4: 293T cell lysate at 10 µg
Predicted band size: 54 kDa
Immunocytochemistry/ Immunofluorescence - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labeling NRF1 with unpurified ab175932 at a dilution of 1/50.
Flow Cytometry (Intracellular) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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.
ChIC/CUT&RUN sequencing - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
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 Rabbit IgG, monoclonal [EPR25A] - Isotype 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 found in the Protocol booklet in the Support and downloads section.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.