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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| M, H, Vrt, R, Pl | WB, IF, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-650 |
| Brand Family | Upstate |
| Trade Name |
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| Description | ChIPAb+ Ubiquityl-Histone H2B - 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+ Ubiquityl-Histone H2B, (clone 56) set includes the Ubiquityl-Histone H2B, (clone 56) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 213 bp region within the coding region of the human GAPDH gene. The ubiquityl-Histone H2B and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Ubiquityl-Histone H2B associated chromatin. |
| Alternate Names |
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| Product Information | |
|---|---|
| Format | Protein G Purified |
| Control |
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| Presentation | Anti-ubiquityl-Histone H2B, clone 56 (mouse monoclonal IgG). One vial containing 25 μg of protein G purified antibody in 25 μg in 50 μL PBS containing 0.05% sodium azide. Store at -20°C. Normal Mouse IgG. One vial containing 25 μg purified mouse IgG in 25 μL storage buffer containing 0.1% sodium azide. Store at -20°C. ChIP Primers GAPDH Coding Region. One vial containing 75 μL of 5 μM of each primer specific for the coding region of human GAPDH. Store at -20°C. FOR: GGC TCC CAC CTT TCT CAT CC REV: GGC CAT CCA CAG TCT TCT GG |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This ChIPAb+ Ubiquityl-Histone H2B -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
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| Application Notes | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa S3 cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 μg of either a normal mouse IgG or Anti-Ubiquityl-Histone H2B antibody and the Magna ChIP G Kit (Cat. # 17-611). Successful immunoprecipitation of ubiquityl-Histone H2B associated DNA fragments was verified by qPCR using GAPDH promoter (negative) and GAPDH coding (positive) Primers (Please see figures). 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 G ChIP™ (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. Western Blot Analysis: HeLa cell lysate was resolved by electrophoresis, transferred to PVDF and probed with anti-Ubiquityl-histone H2B at a dilution of 1:4000 (Please see figures). |
| Biological Information | |
|---|---|
| Immunogen | Immunogen used was a branched peptide corresponding to the ubiquitination site on human histone H2B. |
| Epitope | Lys120 ubiquitination site |
| Clone | Clone 56 |
| Host | Mouse |
| Specificity | Recognizes histone H2B ubiquitinated on lysine 120 but not free ubiquitin or histone H2B in its unmodified state. |
| Isotype | IgG2aκ |
| Species Reactivity |
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| Species Reactivity Note | Human, mouse, rat. Not tested in other species. The immunogen sequence is identical in a wide range of animal and plant species, so broad cross-reactivity is expected. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene encodes a member of the histone H2B family, and generates two transcripts through the use of the conserved stem-loop termination motif, and the polyA addition motif. [provided by RefSeq] |
| Gene Symbol |
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| Modifications |
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| UniProt Number |
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| UniProt Summary | FUNCTION: Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Has broad antibacterial activity. May contribute to the formation of the functional antimicrobial barrier of the colonic epithelium, and to the bactericidal activity of amniotic fluid. SUBUNIT STRUCTURE: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. SUBCELLULAR LOCATION: Nucleus. PTM: Monoubiquitination of Lys-120 by the RNF20/40 complex gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 Lys-4 and Lys-79 methylation. It also functions cooperatively with the FACT dimer to stimulate elongation by RNA polymerase II. Phosphorylated on Ser-15 by STK4/MST1 during apoptosis; which facilitates apoptotic chromatin condensation. Also phosphorylated on Ser-15 in response to DNA double strand breaks (DSBs), and in correlation with somatic hypermutation and immunoglobulin class-switch recombination. SEQUENCE SIMILARITY: Belongs to the histone H2B family. |
| Molecular Weight | ~25 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa S3 cells (1X 106 cell equivalents per IP) were subjected to chromatin immuno-precipitation using 1 µg of either a normal mouse IgG or Anti-ubiquityl-Histone H2B, clone 56 antibody and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of ubiquityl-Histone H2B-associated DNA fragments was verified by qPCR using Control Primers. Please refer to the EZ-Magna ChIP™ G (Cat. # 17-409) 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. |
| Packaging Information | |
|---|---|
| Material Size | 25 assays |
| Material Package | 25 assays per set. Recommended use: ~1 μg antibody per chromatin immunoprecipitation (dependent upon biological context). |