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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H | WB, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10263 |
| Trade Name |
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| Description | ChIPAb+™ JMJD6 - ChIP Validated Antibody and Primer Set |
| Overview | All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction. The ChIPAb+JMJD1C set includes the JMJD1C antibody, a Normal Rabbit IgG, and control primers which amplify a 110 bp region of ChIP Primers, human β-globin. The JMJD1C and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of JMJD1C -associated chromatin. |
| Alternate Names |
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| Background Information | JmjC domain-containing protein 6 (JMJD6) is a dioxygenase belonging to group of proteins known as the Jmjc-domain containing proteins. This family of proteins is a group of redox enzymes specifically involved in catalyzing oxidation reactions, and are characterized by the presence of a highly conserved jumonji C (Jmjc)-domain. JMJD6 can function both as a lysyl-hydroxylase and a histone arginine demethylase. As a lysyl-hydroxylase, JMJD6 catalyzes 5-hydroxylation of target protein lysine residues. As an arginine demethylase, JMJD6 has a critical part to play in histone code by demethylating of histone H3 and histone H4. It has also been shown to mediate U2AF2/U2AF65 5-hydrozylation, thereby functioning as a regulator for RNA splicing through affecting U2AF2/U2AF65 pre-mRNA activity. JMJD6 is a critical component of embryogenesis for the differentiation of various organs, and may act as a regulator for hematopoietic regulation and macrophage cytokine response. High expression of JMJD6 has been observed in heart, kidney and skeletal muscle. Medium to low expression has been seen in brain, liver, lung, ovary, pancreas, placenta, prostate, testis, thymus, and spleen. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
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| Presentation | Anti-JMJD6 (rabbit polyclonal). One vial containing 50 µL of purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl and 0.05% sodium azide before the addition of glycerol to 30%. Store at -20°C. Concentration: 0.7 mg/mL Normal Rabbit IgG. One vial containing 125 µg of Rabbit IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C. ChIP Primers, human β-globin. One vial containing 75 μL of each primer (5 μM) specific for the human β-globin promoter. Store at -20°C. FOR: AGG ACA GGT ACG GCT GTC ATC REV: TTT ATG CCC AGC CCT GGC TC |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This ChIPAb+ JMJD6 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
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| Application Notes | Chromatin Immunoprecipitation: Representative lot data. Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1.4 µg of either Normal Rabbit IgGor Anti-JMJD6 and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of JMJD6 associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin as a positive locus, and β-actin promoter primers as a negative locus. (Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated. Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. Western Blot Analysis: Representative lot data. HeLa nuclear extract was resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-JMJD6 (1 µg/mL). Proteins were visualized using a Donkey Anti-Rabbit IgG conjugated to HRP and chemiluminescence detection system. Arrow indicates JMJD6 (~46 kDa). (Figure 3). Immunocytochemistry Analysis: Representative lot data. Confocal fluorescent analysis of A431, HeLa cells using Anti-JMJD6 (Red). Actin filaments have been labeled with Alexa Fluor® 488 dye- Phalloidin (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus. (Figure 4). |
| Biological Information | |
|---|---|
| Immunogen | Linear peptide corresponding to human JMJD6. |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | This antibody recognizes JMJD6. |
| Species Reactivity |
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| Species Reactivity Note | Demonstrated to react with human. Predicted to react with Xenopus based on 100% sequence homology. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | This gene encodes a nuclear protein with a JmjC domain. JmjC domain-containing proteins are predicted to function as protein hydroxylases or histone demethylases. This protein was first identified as a putative phosphatidylserine receptor involved in phagocytosis of apoptotic cells; however, subsequent studies have indicated that it does not directly function in the clearance of apoptotic cells, and questioned whether it is a true phosphatidylserine receptor. Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq]. |
| Gene Symbol |
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| Purification Method | Affinity Purfied |
| UniProt Number |
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| UniProt Summary | FUNCTION: Dioxygenase that can both act as a histone arginine demethylase and a lysyl-hydroxylase. Arginine demethylase which demethylates histone H3 at Arg-2 (H3R2me) and histone H4 at Arg-3 (H4R3me), thereby playing a central role in histone code. Also acts as a lysyl-hydroxylase that catalyzes 5-hydroxylation on specific lysine residues of target proteins such as U2AF2/U2AF65 and LUC7L2. Acts as a regulator of RNA splicing by mediating 5-hydroxylation of U2AF2/U2AF65, affecting the pre-mRNA splicing activity of U2AF2/U2AF65. Required for differentiation of multiple organs during embryogenesis. Probably acts as a key regulator of hematopoietic differentiation. Seems to be necessary for the regulation of macrophage cytokine responses. COFACTOR: Binds 1 Fe2+ ion per subunit. SUBUNIT STRUCTURE: Interacts with LUC7L2, LUC7L3 and U2AF2/U2AF65. SUBCLELLULAR LOCATION: Nucleus TISSUE SPECIFICTY: Highly expressed in the heart, skeletal muscle and kidney. Expressed at moderate or low level in brain, placenta, lung, liver, pancreas, spleen, thymus, prostate, testis and ovary. Up-regulated in many patients with chronic pancreatitis. Expressed in nursing thymic epithelial cells. INDUCTION: Up-regulated upon cytokine treatment, but not upon TNF-alpha treatment. DOMAIN: The nuclear localization signal motifs are necessary and sufficient to target it into the nucleus. SEUQENCE SIMILARITIES: Belongs to the JMJD6 family. Contains 1 JmjC domain. CAUTION: Was initially thought to constitute the phosphatidylserine receptor, a receptor that mediates recognition of phosphatidylserine, a specific marker only present at the surface of apoptotic cells. Phosphatidylserine receptor probably participates in apoptotic cell phagocytosis. This protein was identified using phage display expressing mAb 217, an antibody that specifically recognizes phosphatidylserine receptor. However, its nuclear localization and the fact that mAb 217 antibody still recognizes the phosphatidylserine receptor in mice lacking JMJD6, strongly suggest that it does not constitute the receptor for phosphatidylserine and is not involved in apoptotic cell removal. SEQUENCE CAUTION: The sequence AAH47003.1 differs from that shown. Reason: Erroneous initiation. The sequence BAA25511.1 differs from that shown. Reason: Erroneous initiation. |
| Molecular Weight | ~46 kDa observed. Uncharacterized crossreacting band observed below the 46 kDa target band in some lysates. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Representative lot data. Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1.4 µg of either Normal Rabbit IgG or Anti-JMJD6 and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of JMJD6 associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin (Figure 1). Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. |
| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage. |
| Packaging Information | |
|---|---|
| Material Size | 25 assays |
| Material Package | 25 assays per set. Recommended use: 1.4 µg (2 µL) of antibody per chromatin immuno-precipitation (dependent upon biological context). |