Homeobox protein Nkx-2.5 Antibodies

Homeobox protein Nkx-2.5 (NKX2-5)

Implicated in commitment to and/or differentiation of the myocardial lineage. Acts as a transcriptional activator of ANF in collaboration with GATA4 (By similarity).

Binds to the core DNA motif of NPPA promoter (PubMed:22849347, PubMed:26926761). It is transcriptionally controlled by PBX1 and acts as a transcriptional repressor of CDKN2B (By similarity).

Gene Information

Human
Gene Name:	NKX2-5
Uniprot:	P52952
Entrez:	1482

Family

Belongs to:
NK-2 homeobox family

Alternative Names

cardiac-specific homeo box; cardiac-specific homeobox 1; Cardiac-specific homeobox; CHNG5; CSX1; CSXFLJ97195; FLJ52202; FLJ99536; Homeobox protein CSX; Homeobox protein NK-2 homolog E; homeobox protein Nkx-2.5; NK2 transcription factor related, locus 5 (Drosophila); NKX2.5; NKX2.5FLJ97166; NKX2-5; NKX2E; NKX2EFLJ97197; NKX4-1; tinman paralog

Molecular Weight

Mass (kDA):
34.918 kDA

Genomic Context

Human
Location:	5q35.1
Sequence:	5; NC_000005.10 (173232109..173235321, complement)

Tissue Specificity

Expressed only in the heart.

Subcellular Localizations

Nucleus.

Diseases

• Congenital Heart Defects
• Heart Diseases
• Congenital Heart Disease
• Heart Septal Defects
• Atrial Septal Defects
• Ventricular Septal Defects
• Congenital Abnormality
• Hypertrophy
• Haploinsufficiency
• Myocardial Infarction
• Tetralogy Of Fallot
• Heart Failure
• Hypoplasia

• Cardiomyopathies
• Infarction
• Cardiac Arrhythmia
• Atrioventricular Block
• Malignant Neoplasms
• Cardiac Hypertrophy
• Heart Block

Interacting Proteins

• Fblim1
• GATA4

Pathways

• Heart Development
• Heart Formation
• Cell Differentiation
• Pathogenesis
• Cardiac Conduction
• Regeneration
• Methylation
• Cell Proliferation
• Localization
• Cell Cycle
• Gastrulation
• Heart Morphogenesis
• Translation

• Dna Methylation
• Aging
• Tube Formation
• Mesoderm Formation
• Nuclear Export
• System Development
• Cell Growth

PTMs

• Methylation
• Phosphorylation
• Acetylation
• Sumoylation
• Ubiquitination

• Glycosylation
• Demethylation
• Phosphorothioation
• Cleavage
• Deacetylation

For details, visit:
bosterbio.com/bosterbio-gene-info-cards/NKX2-5

Best Uses Of The NKX2-5 Marker

This article is about perinatal failing heart and the NKX2-5 marker. It discusses AntiNKX2-5 Marker, a newly developed antibody that recognizes the NKX2-5 molecule. The reporter gene is transactivated by it. In addition, we will discuss the NKX2-5Marker and its use in animal research.

NKX2-5 Marker

The NKX2-5 transcription factor is a redundant transcription element that is necessary to reinduct ANF in perinatally failing hearts. This redundant function is unique to the NKX2-5 marker. This study is the first to use an NKX2-5 gene-targeting construct to identify the alterations in ANF expression in neonatal and adult hearts.

The consensus binding site for Nkx2-5 is located at bp.-260 and relative to the beginning site of ANF transcript. In the presence of ANF, the Nkx2-5 marker binds at this site. Results are normalized to bactin expression. A promoterless reporter is used to measure luciferase activity when the Nkx2-5 gene has been expressed.

The NKX2-5 genome is a candidate gene for Nkx2-5 mRNA expression. It also has a number of other uses. Recent studies have linked the gene to diabetes and cardiovascular disease. The expression of NKX2-5 is essential for the differentiation between Tbx18+ EPCs and pacemaker cells. Bmp4 promotes Gata4, a gene that regulates NKX2.5 expression.

The regulation of ANF genes under many pathological and physiological conditions is possible through the NKX2-5 gene. Tamoxifen injections at E19 result in progressive heart disease within a few months. The gene deletion causes death. Northern blotting detects time-dependent changes of mRNA levels. In both control and Nkx2-5 knockout hearts, mRNA levels were decreased.

NKX2-5 Marker in perinatal failing hearts

We have previously identified a novel candidate NKX2-5 protein as a cause in perinatal failing hearts disease. This protein plays a role in maintaining cardiac mechanics. We have investigated the role of the NKX2-5 Marker in perinatal failing hearts by measuring changes in integrin b-1 levels and connexin 43 in neonatal and adult mouse hearts.

We studied a patient who had a splice-site mutation in the NKX2-5 genes. He was born with a mutation in exon 1 of the NKX2.5 gene. This patient was 12 years of age and presented with recurrent syncope. His heart weight was 300g, and a pacemaker had been installed. The father, aged 29 years old, died suddenly. The autopsy report found no other abnormalities.

Mutations in NKX2-5 can be associated with a variety cardiac anomalies such as atrial fibrillation or ventricular septal defect. These phenotypes have not yet been shown to be universal. It is possible that mutations in NKX2-5 could cause CHD.

The NKX2-5Marker was found to be overexpressed in the fetal cardiac muscle at various stages of development. This gene also mediates the expression of many important genes, including troponins. This gene may help to identify patients suffering from hypertrophic Cardiomyopathy. A large number of studies are being conducted to determine the role that the NKX2-5 marker plays in perinatal failing heart.

This study examined how ECM expression differs between mutant and wild-type mice hearts. In particular, the expression of collagen IV and fibronectin were significantly increased in mutant mouse hearts. However, there were no significant differences between mutant and wild-type hearts in terms of expression of ECM proteins. In addition, we observed an increased expression of connexin 43 in mutant hearts. This protein is essential for maintaining the structural integrity of the heart and regulates cardiac contractility.

Transactivated reporter gene by NKX2-5 Marker

The NKX2-5Marker transcription factor has multiple downstream targets. These targets regulate the development myocardium as well as the endocardium. This protein has been implicated for many human diseases. Its downstream targets include genes involved in the regulation of the redox state of the heart, the NMDA receptor, and endothelial cells. It is not known what exact roles these genes play.

The C-set peaks of NKX2-5 mutant proteins and WT proteins were significantly enriched for motifs of broad range expression TFs. The B-set peaks in the NKX2-5 HD mutation protein were overlapping with 1% of its target gene (NKX2-5YRDY–A). The NKX2-5 marker transactivated a reporter genome. In the C-set, the TF binding sites were tested to determine if there were cofactors that NKX2-5 used to guide it off-targets. TFs specifically involved in E-26 transformation were found to be significantly enriched in the C-set of mutant protein peaks.

Mutants in NKX2-5 can recognize a wide range targets uniquey. These NKX2-5 WT molecules associate with a large C-set of off-targets. CHD signatures can also be determined by the relative C/set size and the B-set. The NKX2-5HD targets include genes related to chromatin organization, the cell cycle, as well as many widely expressed TFs.

DamID was also successful at identifying NKX2-5 WT targets. He et.al. conducted the RNA-Seq experiment, which was mentioned by the authors. We will provide more information on the results. Their data show that NKX2-5 Marker transactivated reporter gene expression levels are highly dependent on the presence of other RNA-binding genes, including genes that are regulated by RNA-Seq.

Anti-NKX2-5 Marker

The Anti NKX2-5 (Boster Bio) Marker is a Rabbit Polyclonal Antibody against the human NKX2protein. This antibody binds with the 19 amino acid synthetic protein near the center human NKX2protein. This antibody was validated through Immunoassay as well as EIA techniques. Boster Bio offers high-affinity and specificity antibodies and ELISA kits that have been cited in over 18,000 publications. Boster offers a quality guarantee and each product has been rigorously tested for various applications. Boster Bio expands its product range on a monthly base.

The heart's SAN regulates the heartbeat, and rhythm. During the SAN's development, many genes are involved. These genes form a genetic web. Shox2 (a short stature homeobox) regulates Nkx2.5 early cardiac differentiation marker. Nkx2-5 expression patterns and Shox2 expression patterns are mutually exempt during the early stages SAN/venous pole development.

Atrioventricular dysfunction can occur in mice and humans that carry the Nkx2-5 mutation. The number of cardiac myocytes in murine's conduction system is affected by the Nkx2-5-gene dosage. Null mutant embryos don't have minK-lacZ expressing cells in the less curvature, which is where the AV Node originates. Haploinsufficient embryos, as well as adults, exhibit cellular hypoplasia both in the central and peripheral conduction system. Cx40 and Cx45 expressions are also associated with the development the AV node.

Nkx2-5 mutant embryos lack both AV node primordium and minK-lacZ activity. They also show abnormal heart tube development and somite staining. These results are consistent for WT embryos. It is necessary to perform a Southern Blot analysis in order to confirm that Nkx2-5 mutations have occurred in a particular embryo. An antibody that recognizes Nkx2-5 protein can also detect Nkx2-5 absence in an embryo.

Amplification of Mouse Hey2 cDNA by PCR

To test gene expression, total heart rate RNA was isolated at 2-day-old -/ littermates. EtBr staining confirmed equal amounts of DNA were loaded into the hybridization processes. The results of hybridization with an ANF probe revealed that Hey2-/ ventricles showed strong induction. Both genotypes show increased expression of ANF. However, the Hey2 knockout mouse shows stronger ventricular expression. The expression of CARP mRNA in Hey2 knockout mice was also higher.

Mice lacking Hey2 exhibit a profound cardiac disorder. Their heart-to-body weight ratios rose by 30%-300% depending on the age. Hey2-/ mice displayed larger cardiomyocytes and enlarged atria. Mice expressing high levels Hey2 also had increased myofibrillar disarray which may have contributed to massive cardiac enlargement.

While Hey1/2 DKO mice are fatal at young ages (see below), conditional inactivation allows us to study the gene's function in adult vascular development. The Hey family bHLH transcription elements are direct targets of Notch and regulate the phenotypes for most vertebrate bodies. Hey1 and Hey2 are both required for vascular development, but neither of them causes overt vascular deficiency.

HUVECs that are Hirsutin-mediated undergo apoptosis. The result is increased expression and speed of H-reactive (H-rhodopsin). EndoMT contributes to radiation proctitis. The incidence of intestinal cancer is also reduced by Hey2 overexpression.