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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, M, R, Rb, Chp, Eq, Po | WB, DB, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10264 |
| Trade Name |
|
| Description | ChIPAb+™ JHDM1B Antibody |
| 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+JHDM1B set includes the JHDM1B antibody, a Normal Rabbit IgG, and control primers which amplify a 110 bp region of ChIP Primers, human β-globin. The JHDM1B and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of JHDM1B -associated chromatin. |
| Alternate Names |
|
| Background Information | JHDM1B (jumonji domain-containing histone demethylation protein 1B) and KDM2B. Due to the presence of a F-box motif, this protein is also known as the F-box/LRR-repeat protein 10 (FBXL 10). This protein functions as a histone demethylase for lysine 36 of histione H3 and preferentially demethylates dimethylated lysine 3 with weak or no activity for mono- and tri-methylted H3 lysine 36. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
|
| Presentation | Components: Anti-JHDM1B (Rabbit Polyclonal). One vial containing 50 µL of purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4) and 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+ JHDM1B -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
|
| 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 2 µL of either Normal Rabbit IgG or Anti-JHDM1B and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of JHDM1B 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 probed with Anti-JHDM1B (1 ug/mL). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates JHDM1B (~165 kDa). (Figure 3). An uncharacterized band appears at ~109 kDa in some lysates. Immunocytochemistry Analysis: Representative lot data. Confocal fluorescent analysis of NIH/3T3 and HeLa cells using Anti-JHDM1B (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 | KLH-conjugated linear peptide corresponding to human JHDM1B. |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Species Reactivity |
|
| Species Reactivity Note | Demonstrated to react with Human and Mouse. Predicted to react with Rat, Rabbit, Chimpanzee, Equine, and Porcine based on 100% sequence homology. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
|
| Entrez Gene Summary | This gene encodes a member of the F-box protein family which is characterized by an approximately 40 amino acid motif, the F-box. The F-box proteins constitute one of the four subunits of ubiquitin protein ligase complex called SCFs (SKP1-cullin-F-box), which function in phosphorylation-dependent ubiquitination. The F-box proteins are divided into 3 classes: Fbws containing WD-40 domains, Fbls containing leucine-rich repeats, and Fbxs containing either different protein-protein interaction modules or no recognizable motifs. The protein encoded by this gene belongs to the Fbls class. Multiple alternatively spliced transcript variants have been found for this gene, but the full-length nature of some variants has not been determined. [provided by RefSeq]. |
| Gene Symbol |
|
| Purification Method | Affinity Purfied |
| UniProt Number |
|
| UniProt Summary | FUNCTION: Histone demethylase that demethylates Lys-4 and Lys-36 of histone H3, thereby playing a central role in histone code. Preferentially demethylates trimethylated H3 Lys-4 and dimethylated H3 Lys-36 residue while it has weak or no activity for mono- and tri-methylated H3 Lys-36. Preferentially binds the transcribed region of ribosomal RNA and represses the transcription of ribosomal RNA genes which inhibits cell growth and proliferation. May also serve as a substrate-recognition component of the SCF (SKP1-CUL1-F-box protein)-type E3 ubiquitin ligase complex. CATALYTIC ACTIVITY: Protein N6,N(6)-dimethyl-L-lysine + 2-oxoglutarate + O2 = protein N(6)-methyl-L-lysine + succinate + formaldehyde + CO2. Protein N(6)-methyl-L-lysine + 2-oxoglutarate + O2 = protein L-lysine + succinate + formaldehyde + CO2. COFACTOR: Binds 1 Fe2+ ion per subunit (By similarity). SUBUNIT STRUCTURE: Directly interacts with SKP1A and CUL1 (By similarity). SUBCELLULAR LOCATION: Nucleus › nucleolus DOMAIN: The JmjC domain mediates demethylation activity (By similarity). It is also required for repression of ribosomal RNA genes. SEQUENCE SIMILARITIES: Belongs to the JHDM1 histone demethylase family. Contains 1 CXXC-type zinc finger. Contains 1 F-box domain. Contains 1 JmjC domain. Contains 4 LRR (leucine-rich) repeats. Contains 1 PHD-type zinc finger. SEQUENCE CAUTION: The sequence AAH08735.2 differs from that shown. Reason: Erroneous initiation. The sequence BAB55112.1 differs from that shown. Reason: Erroneous initiation. The sequence BAB55301.1 differs from that shown. Reason: Erroneous initiation. The sequence BAC11159.1 differs from that shown. Reason: Erroneous initiation. |
| Molecular Weight | ~165 kDa observed. An uncharacterized lower molecular weight band may be observed 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 2 µL of either Normal Rabbit IgG or Anti-JHDM1B and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of JHDM1B 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 | |
|---|---|
| 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). |