NR4A1 (Ab-351) Antibody
Description
Antibody Structure and Function
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Immunogen: Generated from a synthetic phosphopeptide corresponding to amino acids surrounding Ser351 of human NR4A1 .
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Clonality: Polyclonal, purified via protein A and peptide affinity chromatography .
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Reactivity: Primarily recognizes phosphorylated NR4A1 in human, mouse, and rat tissues .
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Applications: Validated for Western blot (WB), immunohistochemistry (IHC), and enzyme-linked immunosorbent assay (ELISA) .
Biological Significance of NR4A1
NR4A1, also known as Nur77, is an orphan nuclear receptor involved in:
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Cancer Progression: Overexpressed in lung, breast, liver, and colon cancers, correlating with poor prognosis .
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Metabolic Regulation: Interacts with AMPK to modulate energy homeostasis .
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Addiction Pathways: Mediates transcriptional changes during cocaine abstinence .
The phosphorylation at Ser351 is a key regulatory site, influencing NR4A1’s transcriptional activity and subcellular localization .
Cancer Studies
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Breast Cancer: Used to detect NR4A1 phosphorylation in MDA-MB-231 cells, linking it to PSPC1 regulation and tumor growth inhibition .
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Lung Cancer: Employed in IHC to study NR4A1’s role in metastatic progression .
Metabolic Regulation
Neurological Disorders
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Cocaine Addiction: Validated for detecting NR4A1 activation in nucleus accumbens (NAc) neurons during abstinence .
Key Research Findings
Product Specs
Target Background
- This study uncovers a unique mechanism for suppressing hepatocellular carcinoma by switching from glycolysis to gluconeogenesis through Nur77 antagonism of PEPCK1 degradation. PMID: 28240261
- Our research demonstrates that the BCR/BTK target gene NR4A1 is a potential oncogene in mantle cell lymphoma. PMID: 29167454
- This study confirms that NR4A1 sensitizes gastric cancer cells to TNFalpha-induced apoptosis through the inhibition of JNK/Parkin-dependent mitophagy. PMID: 29207128
- Inhibition of NR4A1 in stromal cells increased the TGF-beta1-dependent elevated expression of fibrotic markers, and loss of NR4A1 stimulated fibrogenesis in mice with endometriosis. PMID: 29669342
- Data indicates that NR4A1 knockdown partly decreased surface NR2B by promoting NR2B internalization. PMID: 27876882
- Data demonstrates that SUMOylation is crucial in controlling NR4A1 function in inflammatory cytokine signaling and regulating macrophage cell death. PMID: 28622293
- Our findings suggest that hypoxia-induced down-regulation of TR3 might play a significant role in hypoxia-induced apoptosis resistance in NSCLC. PMID: 28838387
- Nur77 suppresses CD4(+) T cell proliferation and reveals a suppressive role for Irf4 in TH2 polarization; halving Irf4 gene-dosage leads to increases in GATA3(+) and IL-4(+) cells. PMID: 28538176
- Our data show that NR4A1 protein physically associates with the WT1 promoter, and enhanced WT1 promoter transactivation and knockdown of WT1 in MIN6 cells induced apoptosis. These findings suggest that NR4A1 protects pancreatic beta-cells against H2O2 mediated apoptosis by up-regulating WT1 expression. PMID: 28342843
- NR4A modulates the decidualization of hESCs by upregulating prolactin (PRL) and insulin-like growth factor binding protein-1 (IGFBP-1) expression and transformation in vitro. PMID: 27515096
- DNMT1 causes NR4A1 DNA hypermethylation and blocks insulin signaling in Chinese patients with type 2 diabetes. PMID: 27322146
- Data shows that nuclear receptor 4A1 (NR4A1) knockdown and the C-DIM/NR4A1 antagonists were comparable as inhibitors of NR4A1-dependent genes/pathways. PMID: 27144436
- mRNAs expression and methylation pattern of RARB, NR4A1 and HSD3B2 genes in human adrenal tissues (HAT) and in pediatric virilizing adrenocortical tumors (VAT) were analyzed. PMID: 27670690
- The NR4A sub-family of nuclear orphan receptors (Nor-1, Nurr-1 and Nur-77) may play a role in trophoblastic cell differentiation. PMID: 28808448
- beta1-integrin expression is regulated in pancreatic and colon cancer cells by the pro-oncogenic orphan nuclear receptor 4A1. PMID: 28418095
- Inhibition of NR4A1-mediated transcriptional activity was involved in the anticancer effects of fangchinoline; fangchinoline represents a novel class of mechanism-based anticancer agents targeting NR4A1 that is overexpressed in pancreatic cancer. PMID: 28754437
- Transcript analysis of four different aggressive lymphoma cell lines overexpressing either NR4A3 or NR4A1 revealed that apoptosis was driven similarly by induction of BAK, Puma, BIK, BIM, BID, and Trail. Overall, our results showed that NR4A3 possesses robust tumor suppressor functions of similar impact to NR4A1 in aggressive lymphomas. PMID: 28249906
- NR4A1 expression is specific to a quiescent subset of T-cells. PMID: 27617863
- We demonstrate that endogenous Nur77 protein expression can serve as a reporter of T-cell receptor and B-cell receptor specific signaling in human peripheral blood mononuclear cells. PMID: 27940659
- We report overexpression of the nuclear receptor NR4A1 in rhabdomyosarcomas that is sufficient to drive high expression of PAX3-FOXO1A. PMID: 27864345
- Study found a marked down-regulated gene expression of the NR4A subfamily (NR4A1, NR4A2, and NR4A3) obtained from Parkinson's disease patients, but only a NR4A1 decrease in Alzheimer's disease patients compared to healthy controls. This study reports that the entire NR4A subfamily and not only NR4A2 could be systemically involved in Parkinson's disease. PMID: 27159982
- Nur77 overexpression prevented pulmonary artery smooth muscle cells from proliferation and migration; Nur77 is an important mediator of hypoxia-induced pulmonary vascular remodeling in pulmonary hypertension. PMID: 27871853
- Data shows Kruppel-like factor 12 (KLF12) impairs endometrial decidualization by transcriptionally repressing Nur77 protein, and Nur77 overexpression reverses the poor decidual response of endometrial stromal cells (hESCs) in recurrent implantation failure (RIF) patients. PMID: 28359310
- Nur77 decreases ET-1 expression by suppressing NF-kappaB and p38 MAPK. PMID: 27765761
- NR4A1 is highly expressed in a subset of HGSOC samples from patients that have a worse progression-free survival. PMID: 26946093
- Nur77 gene expression levels might involve different autonomy of ACTH production between Cushing disease (CD) and subCD. PMID: 27025408
- Data shows that three TR3 orphan nuclear receptor (TR3) transcript variant messenger RNAs (mRNAs) are expressed in human umbilical vein endothelial cell (HUVEC) and are differentially regulated by vascular endothelial growth factor (VEGF). PMID: 26440050
- Mutagenesis of residues lining the identified interaction site on Bcl-B negated the interaction with Nur77 protein in cells and prevented Nur77-mediated modulation of apoptosis and autophagy. PMID: 27129202
- NR4A1 regulates beta1-integrin expression and beta1-integrin-dependent migration of breast cancer cells, and this is accompanied by decreased expression of beta3-integrin. PMID: 26929200
- Data indicate that N-myc downstream regulated gene 1 (NDRG1) competitively bind to glycogen synthase kinase 3beta (GSK-3beta) and orphan nuclear receptor (Nur77) to prevent beta-catenin degradation. PMID: 26359353
- Analysis of amino acid fragments required by TR3/Nur77 for its functions in angiogenesis. PMID: 26155943
- The results demonstrate that Nur77 is induced by oxLDL via the p38 MAPK signaling pathway, which is involved in the regulation of cell survival. Nur77 enhanced cell survival via suppressing apoptosis, without affecting cell proliferation of activated macrophages, which may be beneficial in patients with atherosclerosis. PMID: 26768365
- NR4As regulate gene transcription primarily through interaction with distal enhancers that are co-enriched for NR4A1 and ETS transcription factor motifs. PMID: 26938745
- miR-124 is downregulated in instances of medulloblastoma where Nur77 is upregulated, resulting in a proliferative state that contributes to cancer progression. PMID: 26840408
- ApoA-IV colocalizes with NR4A1, which suppresses G6Pase and PEPCK gene expression at the transcriptional level, reducing hepatic glucose output and lowering blood glucose. PMID: 26556724
- NR4A1 is a key factor in various diseases, including arthritis, inflammation, cancer and cardiovascular diseases. PMID: 25917081
- Identified Nur77/Nor1 as novel regulators of thrombomodulin expression and function in vascular endothelial cells. PMID: 26634653
- The results indicate that genetic variants of the NUR77 gene are associated with an increased risk for both UC and CD. PMID: 26564988
- High NR4A1 expression is associated with breast cancer. PMID: 26229035
- Molecular dynamics simulation results of NR4A1 reveal a pronounced pocket that binds the known ligands, which is similar to known nucleotide-binding sites. Its molecular conformation can be affected by alternate-site modulators. PMID: 26270486
- This study examined the expression and function of TR3 in skin. It also investigated the function of TR3 in the effect of androgens in keratinocytes by treating HaCaT keratinocytes and primary human keratinocytes with dihydrotestosterone (DHT) and testosterone (T). PMID: 26707825
- Nur77 protein is expressed in colon tissues from Crohn's disease and ulcerative colitis. Nur77 also strongly decreased expression of MCP-1, CXCL1, IL-8, MIP-1alpha and TNFalpha in gut epithelial Caco-2 cells. PMID: 26241646
- High NR4A1 expression is associated with Renal Cell Adenocarcinoma. PMID: 26035713
- In hepatocytes, HCV core protein increases drug resistance and inhibits cell apoptosis by inhibiting the expressions of NR4A1 and RUNX3. PMID: 26392314
- Results indicate that Nur77 was overexpressed in a high percentage of human colon and liver cancer, and the intracellular location of Nur77 correlated with elevated serum total Bile acids levels in patients with colon cancer. PMID: 25232032
- A2M is expressed in the vasculature and NR4A receptors modulate VSMC MMP2/9 activity by various mechanisms including the up-regulation of A2M. PMID: 25809189
- This review provides a concise overview of the current understanding of the important metabolic roles governed by NR4A members NR4A1, NR4A2 and NR4A3, including their involvement in a number of diseases. PMID: 25089663
- Data shows that some rexinoids display selective coactivator (CoA) recruitment by the retinoid X receptors (RXRs) homodimer and by the heterodimers nuclear receptor Nur77/RXR and Nurr1/RXR. PMID: 26148973
- Histone acetylation contributes to the regulation of NR4A1 expression in hypercholesterolaemia, and NR4A1 expression reduces hypercholesterolaemia-induced inflammation. PMID: 26396259
- While NR4A1, induced by PDGF-BB, suppresses cell growth on a solid surface, it increases anchorage-independent growth. PMID: 25269081
Q&A
Basic Research Questions
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What is the specificity of NR4A1 (Ab-351) Antibody and how can researchers validate it?
The NR4A1 (Ab-351) Antibody specifically recognizes a region around the phosphorylation site of serine 351 in the human Nuclear Receptor NR4A1 protein. The antibody detects endogenous levels of total Nuclear Receptor NR4A1 protein .
For validation, researchers should:
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Perform western blotting with positive controls (tissues/cells known to express NR4A1, such as human colon carcinoma)
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Include negative controls through knockdown experiments using RNAi against NR4A1
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Verify band size matches predicted molecular weight (approximately 64 kDa)
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Consider parallel staining with another validated NR4A1 antibody targeting a different epitope
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For phosphorylation-specific validation, treat samples with phosphatase inhibitors versus phosphatase treatment
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What experimental applications are validated for NR4A1 (Ab-351) Antibody?
NR4A1 (Ab-351) Antibody has been validated for multiple experimental techniques:
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Immunohistochemistry (IHC) on paraffin-embedded sections, as demonstrated with human colon carcinoma tissue
When designing experiments, researchers should first confirm reactivity with their species of interest (primarily human, though some cross-reactivity with mouse and rat has been reported) and optimize antibody concentration for each specific application and experimental system.
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How should researchers interpret NR4A1 localization patterns in immunostaining experiments?
When using NR4A1 (Ab-351) Antibody for immunostaining:
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In baseline conditions, expect predominantly nuclear localization in most cell types
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During stress responses or specific stimuli, observe for cytoplasmic and potentially mitochondrial translocation
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Phosphorylated NR4A1 (at Ser351) may show distinct localization patterns compared to total NR4A1
For accurate interpretation, researchers should note that NR4A1 can show dynamic localization between the nucleus, cytosol, and mitochondria depending on cellular context . High-throughput single-cell imaging studies with phospho-NR4A1 (ser351) antibodies have confirmed the loss of both nuclear and cytoplasmic NR4A1 under certain conditions , suggesting complex regulation of this protein's localization.
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Advanced Research Questions
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How can NR4A1 (Ab-351) Antibody be used to study NR4A1's role in gene regulation through ChIP assays?
For chromatin immunoprecipitation (ChIP) studies:
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Protocol optimization:
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Use dual crosslinking (DSG followed by formaldehyde) to better capture transcription factor interactions
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Sonicate chromatin to fragments between 200-500bp
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Immunoprecipitate with 5-10μg of NR4A1 (Ab-351) Antibody per 25μg of chromatin
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Target gene selection:
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Data interpretation:
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Research has shown NR4A1 may bind predominantly to gene bodies rather than promoters (<3% promoter localization)
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Evidence suggests NR4A1 binding to IEG genebodies and 3'-UTR overlaps with open chromatin domains identified by ATAC-seq
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Compare NR4A1 binding patterns with other transcription factors like FOS, which exhibits expected localization to promoter regions (42%)
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What experimental approaches allow for studying NR4A1 phosphorylation dynamics in stress responses?
To study NR4A1 phosphorylation during stress responses:
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Stress induction protocols:
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Serum starvation (24 hours) followed by serum replenishment to simulate replication stress
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Compare with other stimuli: DNA damage, heat shock, or specific ligand treatment
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Phosphorylation analysis:
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Subcellular localization tracking:
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Implement time-course immunofluorescence studies following stress induction
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Research has shown that during serum stress, NR4A1 shows dynamic association with the RNA polymerase elongation complex
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Co-immunoprecipitation experiments have demonstrated that RNA Pol II interacts with NR4A1 under baseline conditions, but this association is not detectable upon serum stress
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How should researchers design NR4A1 knockdown/overexpression studies to investigate its functional role?
For functional studies of NR4A1:
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Knockdown approaches:
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Overexpression strategies:
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Functional readouts:
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What are the experimental considerations for using NR4A1 (Ab-351) Antibody in age-related cognitive decline research?
For cognitive decline research:
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Age-dependent analysis:
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Experimental models:
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Mechanistic studies:
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Technical considerations:
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How can researchers use NR4A1 (Ab-351) Antibody to investigate protein-protein interactions in different subcellular compartments?
For protein interaction studies:
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Co-immunoprecipitation approaches:
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Proximity-based methods:
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Implement proximity ligation assays (PLA) to visualize NR4A1 interactions in situ
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Use BioID or APEX2 proximity labeling with NR4A1 fusions to identify novel interaction partners
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Subcellular distribution analysis:
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Conduct subcellular fractionation followed by immunoblotting with NR4A1 (Ab-351) Antibody
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NR4A1 shows complex distribution patterns including nuclear export to the cytosol (XPO1-mediated)
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Translocation to mitochondria has been observed upon interaction with RXRA and in the presence of 9-cis retinoic acid
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Monitor kinase STK11 sequestration by NR4A1 in the nucleus, which attenuates cytoplasmic AMPK activation
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