KDM6B Antibodies

Lysine-specific demethylase 6B (KDM6B)

Histone demethylase that especially demethylates'Lys- 27' of histone H3, thereby playing a central role in histone code (PubMed:17825402, PubMed:17851529, PubMed:17713478, PubMed:18003914). Demethylates trimethylated and dimethylated H3 'Lys-27' (PubMed:17825402, PubMed:17851529, PubMed:17713478, PubMed:18003914).

Plays a central role in regulation of posterior development, by regulating HOX gene expression (PubMed:17851529). Involved in inflammatory reaction by engaging in macrophage differentiation in case of inflammation by regulating gene expression and macrophage differentiation (PubMed:17825402).

Gene Information

Human
Gene Name:	KDM6B
Uniprot:	O15054
Entrez:	23135

Family

Belongs to:
UTX family

Alternative Names

EC 1.14.11; EC 1.14.11.-; JmjC domain-containing protein 3; JMJD3; JMJD3lysine-specific demethylase 6B; jumonji domain containing 3, histone lysine demethylase; Jumonji domain-containing protein 3; KDM6B; KIAA0346jumonji domain containing 3; Lysine (K)specific Demethylase 6B; Lysine (K)-specific Demethylase 6B; Lysine demethylase 6B

Molecular Weight

Mass (kDA):
176.632 kDA

Genomic Context

Human
Location:	17p13.1
Sequence:	17; NC_000017.11 (7834210..7854796)

Subcellular Localizations

Nucleus.

Diseases

• Malignant Neoplasms
• Neoplasms
• Inflammation
• Retinoblastoma
• Leukemia
• Lymphoma
• Prostatic Neoplasms
• Teratoma
• Infective Disorder
• Malignant Neoplasm Of Prostate
• Neoplasm Metastasis
• Malignant Neoplasm Of Breast
• Carcinoma

• Acute Promyelocytic Leukemia
• Colonic Neoplasms
• Inflammatory Response
• Congenital Heart Defects
• Radiation Injury
• Autoimmunity
• Malignant Tumor Of Colon

Interacting Proteins

• ESR1

Pathways

• Methylation
• Demethylation
• Gene Silencing
• Neurogenesis
• Senescence
• Cell Differentiation
• Histone Methylation
• Histone Modification
• Cellular Senescence
• Histone Acetylation
• Pathogenesis
• Cell Death
• Neural Tube Closure

• Tissue Development
• Chromatin Remodeling
• Cell Proliferation
• Dna Methylation
• Tube Closure
• Aging
• Stem Cell Maintenance

PTMs

• Methylation
• Demethylation
• Reduction

• Acetylation
• Phosphorylation
• Deacetylation

• Ubiquitination

Research Areas

• Cancer
• Chromatin Research
• Transcription Factors and Regulators

For details, visit:
bosterbio.com/bosterbio-gene-info-cards/KDM6B

Best Uses Of The KDM6B Marker

The KDM6B marker is a inhibitor of synthesis of lipids. It interacts with SNAI1 to reduce the production of lipids. Boster Bio has a unique application for the KDM6B Marker: scientists can submit their findings to obtain product credits, receive product credits and submit specific specimens or species to be tested. Scientists around the world can utilize this product.

qPCR detection of binding of KDM6B to SNAI1 Promoter

In this study, we utilized QPCR to identify KDM6B binding to SNAI1 promoter. To identify KDM6B binding sites, we used primers that target the SNAI1 promoter. For internal control the products were normalized to GAPDH. RQ = 2-DDCt was used to calculate the relative expression level.

KDM6B, ISL1 and SNAI1 have synergistic effects on SNAI1's expression. Together they regulate the demethylation of SNAI1 promoter. This study suggests that ISL1 and KDM6B could hinder the production of fatty acids and increase their degradation. In addition, these proteins can promote autophagy and reduce NAFLD.

We then sought to determine if KDM6B inhibited lung metastasis. In vivo tests showed that the KDM6B mutation in OS cells blocks lung metastasis. We utilized an orthotopic injection model and in vitro transwell to test the effects of KDM6B on OS tumor stem cells. We removed lung tissue from KDM6B-KO mice on day 23 to determine whether KDM6B inhibits lung metastaticsis. After fixing the lungs using Bouin's solution, we stained them with H&E stain. Manually counting metastasis nodules were carried out.

Furthermore, we utilized qPCR to determine whether KDM6B binds to SNAI1 promoter in OS cells. We also examined LDHA, VEGFA and ISL1 mRNA expressions in HOS cells as well as control/KDM6B -KD 143B cells. Moreover, our results indicate an immediate link between KDM6B and the SNAI1 gene in NAFLD.

We also utilized an antibody that modifies H3K27me3 to analyze tumor samples. Our results indicated that KDM6B was associated with lower overall survival among patients with metastatic OS. We also hypothesized that high KDM6B expression was a significant predictor of OS and could be the cause of OS metastasis. We will now explore new ways to study KDM6B more closely.

Moreover, this new technique also uncovers new TGF-b-regulated genes. We will now study their functions and find out the mechanisms involved. Our work emphasizes the need to continue study into the mechanisms that control these genes through TGF-b. These results are encouraging. We are looking forward to investigating the importance of these newly identified genes in the prediction of cancer's prognosis.

Immunofluorescence detection of the KDM6B Marker

Utilizing the Beyotime Biotechnology immunofluorescence kit to KDM6B Marker detection, scientists can determine if the protein is present in living cells. Antibodies against ISL1 and KDM6B are purchased from Cell Signaling Technology and Abcam and Abcam, respectively. The results were viewed under an confocal microscope. The DNA of the nuclear cell was labeled by using 4',6-phenylindole.

This Boster Bio Immunofluorescence detector kit for KDM6B is designed to detect KDM6B binding in the promoter region SNAI1, a key gene involved in the liver disease. KDM6B antibody recognizes this specific protein by measuring its expression qPCR using specific primers to detect KDM6B binding. KDM6B is a well-known upstream factor for SNAI1 and its demethylation has been implicated in metabolic disorders.

KDM6B, HIF-1a are expressed in ESCC cells at a significantly higher level than in normal esophageal tissues. These proteins are co-localized with HIF-1a, CAIX, and were significantly elevated in tumors. KDM6B however did not correlate well with hypoxic markers. Despite these findings , this kit is still useful for finding KDM6B.

The procedure for detecting KDM6B was a modified version of Boster Bio's immunohistochemistry. First cells were fixed with formaldehyde for 10 minutes and then were lysed using 25 mL sepharose protein A. After incubating at 37 degrees Celsius overnight the samples were then centrifuged at 12,000rpm for 5 minutes to separate the DNA/protein complex. The samples were then exposed to light and then treated with phenol/chloroform.

After the immunocytochemistry process after which cells were scraped, they were incubated with the primary antibody (sc-2357) and the secondary antibody (sc-2005) at 4 degC for a period of time. The cells were then subjected to Western Blot (WB), as described previously53. After that, secondary antibodies were added overnight at temperatures of 4 degC and incubated for 1 hour R/T. After that, chemiluminescence could be detected by Clarity WB ECL substrate and ImageJ software.

ISL1 and KDM6B interact to reduce the production of lipids

We have demonstrated that ISL1 is involved with KDM6B in NAFLD. We also discovered that KDM6B/ISL1 was positively related to SNAI1, suggesting that the ISL1/KDM6B may play an important role in the development of NAFLD. These findings suggest that ISL1 interacts with KDM6B in order to inhibit the synthesis of lipids. It could also be a therapeutic target.

In a previous study we found that ISL1 interacts with KDM6 and hinders lipid synthesis. Additionally, we found that SNAI1 expression was reduced by knocking down KDM6B and expressing ISL1. However knocking down KDM6B and overexpressing ISL1 did not inhibit steatosis. These results indicate that lipolysis is dependent on the SNAI1 gene. The interaction between KDM6B and ISL1 could be vital in controlling the synthesis of lipids.

These results support the notion that ISL1 interacts with KDM6b, reducing the production of lipids in NAFLD. ISL1 reduces SNAI1 expression in hepG2 cell cells, which is anti-lipogenic in nature. However, we are yet to investigate whether ISL1 is a liver-specific molecule. These findings are consistent with NAFLD in mice.

Lipogenesis in NAFLD mouse models is inhibited by the overexpression of KDM6B and ISL1. The knockdown of SNAI1 completely eliminates the therapeutic effect. Western Blot analysis revealed that knocking down ISL1 decreased c-caspase 3 as well as the expression of c-caspase 7 genes in NAFLD mice. These results suggest that the knockdown of ISL1 may alter the expression of other lipid-related gene.

In addition to interacting with KDM6B, ISL1 has also been linked to the jumonji domain-containing protein KDM6B. KDM6B is crucial for glucose homeostasis as well as lipid oxidation. ISL1 deletions inhibit this gene's activity. In addition, the overexpression of ISL1 blocks lipid synthesis, while increasing the glucose levels.

Fasting-induced JMJD3 enhances fat acid b-oxidation in human pancreatic and adipose-adipose ad. It is yet to be proven that ISL1 reduces the synthesis of lipids via its own. The inverse relationship may be explained by JMJD3 that is fast-induced. It is a catalyst for fatty acid bi-oxidation.