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17-650 ChIPAb+ Ubiquityl-Histone H2B - ChIP Validated Antibody and Primer Set

17-650
25 assays  25 assays per set. Recommended use: ~1 μg antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      M, H, Vrt, R, PlWB, IF, ChIP
      Description
      Catalogue Number17-650
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ Ubiquityl-Histone H2B - ChIP Validated Antibody and Primer Set
      OverviewAll 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
      • H2BUb
      • Histone H2B (ubiquityl)
      • H2B histone family, member L
      • H2B.1 A
      • histone 1
      • H2bc
      • histone cluster 1
      • H2bc
      References
      Product Information
      FormatProtein G Purified
      Control
      • Includes negative control mouse IgG antibody and primers specific for human GAPDH coding region.
      PresentationAnti-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 LevelMQ100
      Applications
      ApplicationThis ChIPAb+ Ubiquityl-Histone H2B -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Western Blotting
      • Immunofluorescence
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin 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
      ImmunogenImmunogen used was a branched peptide corresponding to the ubiquitination site on human histone H2B.
      EpitopeLys120 ubiquitination site
      CloneClone 56
      HostMouse
      SpecificityRecognizes histone H2B ubiquitinated on lysine 120 but not free ubiquitin or histone H2B in its unmodified state.
      IsotypeIgG2aκ
      Species Reactivity
      • Mouse
      • Human
      • Vertebrates
      • Rat
      • Green Plants
      Species Reactivity NoteHuman, 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 TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryHistones 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
      • H2B.1
      • H2B.a/g/h/k/l
      • H2B/a
      • H2B/g
      • H2B/h
      • H2B/k
      • H2B/l
      • H2BFA
      • H2BFG
      • H2BFH
      • H2BFK
      • H2BFL
      Modifications
      • Ubiquitylation
      UniProt Number
      UniProt SummaryFUNCTION: 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
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin 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
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable 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 Size25 assays
      Material Package25 assays per set. Recommended use: ~1 μg antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+ Ubiquityl-Histone H2B - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ Ubiquityl-Histone H2B 2464584
      ChIPAb+ Ubiquityl-Histone H2B, clone 56 - 2142718 2142718
      ChIPAb+ Ubiquityl-Histone H2B, clone 56 3077132
      ChIPAb+ Ubiquityl-Histone H2B, clone 56 - 2030045 2030045
      ChIPAb+ Ubiquityl-Histone H2B, clone 56 - 2524664 2524664
      ChIPAb+ Ubiquityl-Histone H2B, clone 56 - 3185810 3185810
      ChIPAb+ Ubiquityl-Histone H2B, clone 56 - 3386040 3386040
      ChIPAb+ Ubiquityl-Histone H2B, clone 56 - JBC1787327 JBC1787327
      ChIPAb+ Ubiquityl-Histone H2B, clone 56 - NG1606331 NG1606331
      ChIPAb+ Ubiquityl-Histone H2B, clone 56 - NG1806850 NG1806850

      References

      Reference overviewPub Med ID
      RE-IIBP Methylates H3K79 and Induces MEIS1-mediated Apoptosis via H2BK120 Ubiquitination by RNF20.
      Woo Park, J; Kim, KB; Kim, JY; Chae, YC; Jeong, OS; Seo, SB
      Scientific reports  5  12485  2015

      Show Abstract
      26206755 26206755
      H2B monoubiquitylation is a 5'-enriched active transcription mark and correlates with exon-intron structure in human cells.
      Jung, I; Kim, SK; Kim, M; Han, YM; Kim, YS; Kim, D; Lee, D
      Genome research  22  1026-35  2012

      Show Abstract
      22421545 22421545
      Reelin is a target of polyglutamine expanded ataxin-7 in human spinocerebellar ataxia type 7 (SCA7) astrocytes.
      McCullough, SD; Xu, X; Dent, SY; Bekiranov, S; Roeder, RG; Grant, PA
      Proceedings of the National Academy of Sciences of the United States of America  109  21319-24  2012

      Show Abstract
      23236151 23236151
      Histone crosstalk directed by H2B ubiquitination is required for chromatin boundary integrity.
      Ma, MK; Heath, C; Hair, A; West, AG
      PLoS genetics  7  e1002175  2011

      Show Abstract
      21811414 21811414
      Elongin B-mediated epigenetic alteration of viral chromatin correlates with efficient human cytomegalovirus gene expression and replication.
      Hwang, J; Saffert, RT; Kalejta, RF
      mBio  2  e00023-11  2011

      Show Abstract Full Text Article
      21447700 21447700

      Brochure

      Title
      Advance your Epigenetics Research

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      Product Families

      Categories

      Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies
      Life Science Research > Antibodies and Assays > Primary Antibodies
      Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies