ARNT/HIF-1 beta Antibody Pack

Western Blot: ARNT/HIF-1 beta Antibody Pack [NB100-982] - Analysis of ARNT/HIF-1 beta using ARNT/HIF-1 beta antibody [NB100-110] from ARNT/HIF-1 beta antibody pack [NB100-982] in MCF7 whole cell lysate. Theoretical molecular weight of ARNT/HIF-1 beta is 86.6 kDa, observed molecular weight ~90 kDa.

Immunohistochemistry: ARNT/HIF-1 beta Antibody Pack [NB100-982] - Staining of skin, epidermis using ARNT/HIF-1 beta antibody [NB100-110] from the ARNT/HIF-1 beta antibody pack [NB100-982].

Chromatin Immunoprecipitation: ARNT/HIF-1 beta Antibody Pack [NB100-982] - Allele specific HIF-binding at the rs12814794 associated enhancer. B) Allele-specific qPCR for rs12814794 from DNA fragments isolated in HIF-ChIP experiments (HIF-1A and ARNT/HIF-1 beta) or input DNA from primary renal tubular cells (PTC, n = 4 individuals). DNA from individuals homozygous for the AA or GG genotype were used as positive controls. C) Quantification of the two different alleles (A or G) at rs12814794 in the ChIP-seq reads from HIF-1A and ARNT/HIF-1 beta antibody [NB100-110] immunoprecipitations (PTC #1 Fig 3). Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pgen.1006872) licensed under a CC-BY license.

Western Blot: ARNT/HIF-1 beta Antibody Pack [NB100-982] - Expression of CD44 mRNA in genetically engineered MDA-MB-231 cells. (A) Immunoblot analysis of HIF-1a or HIF-2a expression in whole cell extracts from MDA-MB-231 cells stably expressing EV, HIF-1a-shRNA or HIF-2a-shRNA under normoxia or in response to 4 h treatment with 200 uM CoCl2. ARNT/HIF-1 beta expression was used as a loading control. Theoretical molecular weight of ARNT/HIF-1 beta is 86.6 kDa. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0044078) licensed under a CC-BY license.

Western Blot: ARNT/HIF-1 beta Antibody Pack [NB100-982] - HIF was not the only factor stabilizing activated EGFR in VHL-deficient ccRCC cells. A. Western blot analysis of 786-VHL and 786-mock cells stably expressing shRNA constructs. For HIF2A [NB100-480] and ARNT/HIF-1 beta [NB100-124] analysis, nuclear extracts were generated and analyzed. Anti-HA blot detected HA-VHL. Means and SDs of three separate experiments were shown. Theoretical molecular weight of ARNT/HIF-1 beta is 86.6 kDa, observed molecular weight ~90 kDa. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0023936) licensed under a CC-BY license.

Immunohistochemistry: ARNT/HIF-1 beta Antibody Pack [NB100-982] - Staining of human glioblastoma multi-forme utilizing the ARNT/HIF-1 beta antibody[NB100-124] from the ARNT/HIF-1 beta antibody pack [NB100-982].

• Reactivity: Hu, Mu, Rt, Bv, Ma, Sh
• Applications: WB, ICC/IF, IHC, IP, WB, ChIP

ARNT/HIF-1 beta Antibody Pack Summary

• Description: This pack contains 1 vial of each: NB100-110 (0.1 mL) and NB100-124 (0.1 mL).
• Immunogen: See individual datasheets.
• Specificity: See individual datasheets for specificity notes regarding ARNT/HIF-1 beta antibody pack..
• Gene: ARNT

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP)
  • Immunoblotting
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation
  • Western Blot
• Application Notes: See individual datasheets of components for their validated applications

Reactivity Notes

See individual datasheets of components for their validated species

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.

Alternate Names for ARNT/HIF-1 beta Antibody Pack

• ARNT protein
• ARNT
• aryl hydrocarbon receptor nuclear translocator
• BHLHE2
• Class E Basic Helix-Loop-Helix Protein 2
• Dioxin Receptor, Nuclear Translocator
• HIF1 beta
• HIF-1 beta
• HIF1B
• HIF1BETA
• HIF-1beta
• HIF-1-beta
• HIF1-beta
• hypoxia-inducible factor 1, beta subunit
• Hypoxia-inducible factor 1-beta
• TANGO

Background

Aryl hydrocarbon nuclear translocator (ARNT), also commonly known as Hypoxia-inducible factor 1-beta (HIF-1 beta), is a ubiquitously expressed transcription factor that is part of the basic-helix-loop-helix (bHLH)-Per-ARNT-Sim (PAS) family (1, 2). Human arnt is located on chromosome 1q21 and encodes a protein 789 amino acids (aa) in length with a theoretical molecular weight of 87 kDa (1). Structurally, ARNT has a DNA binding bHLH domain, two PAS domains required for dimerization, and a transactivation domain/PAC region (1). ARNT belongs to the Class II bHLH-PAS proteins and is able to homodimerize or heterodimerize with the Class I proteins including AHA, AHRR, HIF-1 alpha, HIF-2 alpha, NPAS1, and SIM1 (2). Dimerization allows for efficient DNA binding and regulation of their target genes (2).

ARNT has an important role in two specific signaling pathways - the aryl hydrocarbon receptor (AhR) and the hypoxia inducible factor (HIF) pathway (1). In the AhR pathway, AhR in the cytosol is typically inactive and bound to heat shock protein 90 (hsp90) (3). Upon activation and ligand binding by environmental pollutants such as dioxins, AhR is translocated to the nucleus, dissociates from hsp90, and dimerizes with ARNT, leading to binding to response elements and expression of target genes including monooxygenases (1, 3). In the HIF pathway, under hypoxia (low oxygen) conditions prolylhydroxylase domain (PHD) enzymes and factor inhibiting HIF (FIH) are inhibited. HIF-1 alpha (or HIF-2 alpha) accumulates and is transported to the nucleus where it heterodimerizes with ARNT, allowing for binding to target gene's hypoxia response element (HRE), recruitment of coactivators, and transcription (1, 3). HIF-induced gene transcription plays a large role in tumor progression by promoting invasion, metastasis, de-differentiation and altered metabolism, and angiogenesis (1). While HIF-1 alpha's stability is dependent upon oxygen conditions, HIF-1 beta is stable in both normoxia and hypoxia (1-3).

The bHLH-PAS family and ARNT have been linked with a variety of pathologies and diseases including cancer, metabolic diseases, autoimmune diseases, and psychiatric disorders (2). ARNT/AHR is expressed in the skin and its pathway activation enhances skin barrier function and epidermal terminal differentiation, thus AHR agonists are currently being used as therapeutics for atopic dermatitis and psoriasis (4). Accordingly, studies of Arnt-deficient mice show profound abnormalities in skin barrier function and keratinization (4). Additionally, studies suggest that ARNT plays an important role in diabetes and beta-cell function (5). Islets from patients with type 2 diabetes have a significantly decreased ARNT expression compared to glucose-tolerant control donors (5). Modulation and stimulation of the HIF pathway may be a potential therapeutic strategy for treating type 2 diabetes and metabolic syndrome (5).

Alternate names for ARNT/HIF-1 beta include aryl hydrocarbon receptor nuclear translocator, BHLHE2, class E basic helix-loop-helix protein 2, Dixon receptor nuclear translocator, Hypoxia-inducible factor 1-beta, nuclear translocator, and TANGO.

References

1. Mandl, M., & Depping, R. (2014). Hypoxia-inducible aryl hydrocarbon receptor nuclear translocator (ARNT) (HIF-1beta): is it a rare exception?. Molecular medicine (Cambridge, Mass.). https://doi.org/10.2119/molmed.2014.00032

2. Wu, D., & Rastinejad, F. (2017). Structural characterization of mammalian bHLH-PAS transcription factors. Current opinion in structural biology. https://doi.org/10.1016/j.sbi.2016.09.011

3. Esser, C., & Rannug, A. (2015). The aryl hydrocarbon receptor in barrier organ physiology, immunology, and toxicology. Pharmacological reviews.https://doi.org/10.1124/pr.114.009001

4. Furue, M., Hashimoto-Hachiya, A., & Tsuji, G. (2019). Aryl Hydrocarbon Receptor in Atopic Dermatitis and Psoriasis. International journal of molecular sciences. https://doi.org/10.3390/ijms20215424

5. Girgis, C. M., Cheng, K., Scott, C. H., & Gunton, J. E. (2012). Novel links between HIFs, type 2 diabetes, and metabolic syndrome. Trends in endocrinology and metabolism: TEM, https://doi.org/10.1016/j.tem.2012.05.003

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Antibody Packs are guaranteed for 1 year from date of receipt.

Publications for ARNT/HIF-1 beta Antibody Pack (NB100-982)(4)

Publications using NB100-982

• Selfridge AC, Cavadas MA, Scholz CC et al. Hypercapnia Suppresses the HIF-dependent Adaptive Response to Hypoxia. J. Biol. Chem. 2016-05-27 [PMID: 27044749] (WB, Human)
• Fukuda R, Zhang H, Kim JW et al. HIF-1 regulates cytochrome oxidase subunits to optimize efficiency of respiration in hypoxic cells. Cell 2007-04-06 [PMID: 17418790]
• Creighton-Gutteridge, M et al. Cell type-specific, topoisomerase II-dependent inhibition of hypoxia-inducible factor-1alpha protein accumulation by NSC 644221. Cancer Res; 13: 1010-1018. 2007-01-01 [PMID: 17289897]
• Khan Z, Michalopoulos GK, Stolz DB. Peroxisomal localization of hypoxia-inducible factors and hypoxia-inducible factor regulatory hydroxylases in primary rat hepatocytes exposed to hypoxia-reoxygenation. Am J Pathol 2006-10-01 [PMID: 17003483]

FAQs for ARNT/HIF-1 beta Antibody Pack (NB100-982). (Showing 1 - 4 of 4 FAQs).

1. Is this target appropriate for use in Chromatin Research?
   • Yes; here is a list of research areas that we have deemed appropriate for the target "ARNT/HIF-1 beta": Angiogenesis, Autophagy, Cancer, Cellular Markers, Chromatin Research, HIF Target Genes, Hypoxia, Transcription Factors and Regulators, Cardiovascular Biology, Lipid and Metabolism.
2. For use in Western Blot with HIF-1 beta antibodies, what molecular weight of the band should I expect to see?
   • The theoretical molecular weight determined by our technical team for ARNT/HIF-1 beta antibodies is 86.6 kDa.
3. If this product is used in an application or species as a part of a customer review, will that validate this product in the application/species?
   • If any of our primary antibodes are used in an untested application or species and it is shown to work through images from customer reviews or through publications, this validates the application/species for this product, allowing the tested application/species to fall under our 100% guarantee. Please check out our Innovator's Reward Program if you decide to test a primary antibody with a species or application that is not currently listed. Please note that the Innovator's Reward Program only applies to our primary antibodies.
4. Would you please help confirm the components of NB100-982? If NB100-982 really contains regular sizes of these 2 antibodies (not SS), the bargain seems too great to be true. However, if it is composed of NB100-110SS and NB100-124SS, the product seems too expensive.
   • NB100-982 contains 0.1ml of NB100-110 and 0.1ml of NB100-124.

Bioinformatics

• Gene Symbol: ARNT