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17-10054 ChIPAb+ Histone H2B - ChIP Validated Antibody and Primer Set

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17-10054
25 assays  25 assays per set. Recommended use: ~1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      Ch, Xn, H, Yeast (S. cerevisiae)WB, ChIP
      Description
      Catalogue Number17-10054
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ 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+ Histone H2B set includes the Histone H2B antibody, a negative control rabbit IgG, and qPCR primers which amplify a 110 bp region of human β-globin promoter. The Histone H2B and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H2B-associated chromatin.
      Alternate Names
      • H2B
      • Histone H2B
      • H2B histone family, member Q
      • histone 2, H2be
      • histone cluster 2, H2be
      Background InformationHistone H2B is one of the 5 main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N terminal tail H2A is involved with the structure of the nucleosomes of the 'beads on a string' structure.
      References
      Product Information
      FormatPurified
      Control
      • Includes negative control normal rabbit IgG antibody and primers specific for human β-globin.
      PresentationAnti-Histone H2B (rabbit polyclonal). One vial containing 25 µL of protein A purified rabbit polyclonal in buffer containing 0.02 M phosphate buffer, pH 7.6, 0.25 M NaCl, and 0.1% sodium azide, and 30% glycerol. Store at -20°C.

      Normal Rabbit IgG. One vial containing 125 μg of purified rabbit IgG in 125 μL storage buffer. Store at -20°C.

      ChIP Primers, β-globin. One vial containing 75 μL of 5 μM of each primer 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 LevelMQ100
      Applications
      ApplicationThe ChIPAb+ Histone H2B set includes the Histone H2B antibody, a negative control rabbit IgG & qPCR primers which amplify a 110 bp region of human β-globin promoter.
      Key Applications
      • Western Blotting
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µL of Normal Rabbit IgG or 1 µL of Anti-Histone H2B and the Magna ChIP™ A Kit (Cat. # 17-610).
      Successful immunoprecipitation of Histone H2B associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin for a positive, and human GAPDH primers as a negative assay (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 ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

      Western Blot Analysis:
      Representative lot data.
      Recombinant Xenopus Histone H2B, chicken core histones and acid extract from either sodium butyrate or colcemid treated HeLa cells were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Histone H2B (1:5,000 dilution).
      Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).
      Biological Information
      ImmunogenKLH-conjugated, synthetic peptide corresponding to amino acids 118-126 (CG-AVTKYTSSK) of human Histone H2B, with an N-terminal CG added for conjugation purposes.
      HostRabbit
      SpecificityRecognizes and is specific for Histone H2B, MW ~15 kDa. An additional band at ~23 kDa is detected in some samples, which likely corresponds to ubiquityl-Histone H2B.
      Species Reactivity
      • Chicken
      • Xenopus
      • Human
      • Yeast (S. cerevisiae)
      Species Reactivity NoteHuman, chicken, yeast and Xenopus. Others not tested. Broad species cross-reactivity expected based on sequence homology.
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryHistones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H2B family. Transcripts from this gene contain a palindromic termination element.
      Gene Symbol
      • GL105
      • H2B
      • H2B-GL105
      • H2B.1
      • H2B.q
      • H2B/q
      • H2BFQ
      • MGC119802
      • MGC119804
      • MGC129733
      • MGC129734
      • OTTHUMP00000013920
      Purification MethodProtein A Purfied
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: Q8N257 # 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.
      SIZE: 126 amino acids; 13908 Da
      SUBUNIT: 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-121 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.
      SIMILARITY: SwissProt: Q8N257 ## Belongs to the histone H2B family.
      Molecular Weight~15 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 cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µL of Normal Rabbit IgG or 1 µL of Anti-Histone H2B and the Magna ChIP™ A Kit (Cat. # 17-610).
      Successful immunoprecipitation of Histone H2B associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin (Please see figures).

      Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) 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. Note: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
      Packaging Information
      Material Size25 assays
      Material Package25 assays per set. Recommended use: ~1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+ Histone H2B - ChIP Validated Antibody and Primer Set SDS

      Title

      Safety Data Sheet (SDS) 

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

      TitleLot Number
      ChIPAb+ Histone H2B - 1746331C 1746331C
      ChIPAb+ Histone H2B - 2109554 2109554
      ChIPAb+ Histone H2B - 3185815 3185815
      ChIPAb+ Histone H2B - NGR1746331 NGR1746331
      ChIPAb+ Histone H2B - NRG1746331A NRG1746331A
      ChIPAb+ Histone H2B - NRG1746331B NRG1746331B
      ChIPAb+ Histone H2B -2512301 2512301

      References

      Reference overviewPub Med ID
      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
      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 (MilliporeSigma)

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      Product Families

      Categories

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