17-10048 ChIPAb+ Histone H2A.Z - ChIP Validated Antibody and Primer Set

17-10048
25 assays  25 assays per set. Recommended use: ~1 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Übersicht

      Replacement Information

      Key Spec Table

      Species ReactivityKey Applications
      H ChIP, WB
      Description
      Catalogue Number17-10048
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ Histone H2A.Z - 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 H2A.Z set includes the Histone H2A.Z antibody, a negative control antibody (normal sheep IgG), and qPCR primers which amplify a 166 bp region of human GAPDH. The Histone H2A.Z and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H2A.Z -associated chromatin.
      Alternate Names
      • H2A histone family, member Z
      • H2AZ histone
      Background InformationH2A.Z, also known as H2AFZ, is a variant of histone H2A variant that is highly conserved from yeast to humans. Like other histone variants, H2A.Z is encoded by a unique gene, resulting in a slightly divergent type of H2A with specialized functions. H2A.Z is involved in chromatin compaction by stabilizing the histone octamer within the nucleosome. It promotes gene expression through the creation of an open domain of chromatin that is more easily transcribed. H2A.Z is nonrandomly distributed throughout the genome, targeted primarily to pericentric heterochromatin, but excluded from the inactive X chromosome and the nucleolus. H2A.Z is associated with both euchromatin and facultative heterochromatin. DNA methylation can influence chromatin structure and effect gene silencing by excluding H2A.Z, and likewise, H2A.Z protects genes from DNA methylation. Thus, H2A.Z plays a role in regulating heterochromatin silencing and is involved in transcriptional control.
      References
      Product Information
      FormatAffinity Purified
      Control
      • Includes negative control sheep IgG antibody and primers specific for human GAPDH.
      PresentationAnti-Histone H2A.Z (Sheep polyclonal IgG). One vial containing 25 μg of affinity purified sheep polyclonal IgG in PBS, pH 7.4 containing 1% BSA with 0.02% sodium azide, before the addition of 30% glycerol. Store at -20°C.

      Normal Sheep IgG. One vial containing 25 µg of protein G purified sheep IgG in 0.02 M potassium phosphate, 0.15 M NaCl, pH 7.2, with 0.01% sodium azide, before the addition of 30% glycerol. Store at -20°C.

      Control Primers. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH. Store at -20°C.
      FOR: TAC TAG CGG TTT TAC GGG CG
      REV: TCG AAC AGG AGG AGC AGA GAG CGA
      Applications
      ApplicationThis ChIPAb+ Histone H2A.Z -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Chromatin Immunoprecipitation (ChIP)
      • Western Blotting
      Application NotesChromatin Immunoprecipitation:
      Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal Sheep IgG or 1 µg of Anti-Histone H2A.Z and the Magna ChIP™ G Kit (Cat. # 17-611).
      Successful immunoprecipitation of Histone H2A.Z associated DNA fragments was verified by qPCR using Control Primers (Part No. 22-004) for a positive, and GAPDH Coding 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
      HeLa lysate was resolved by electrophoresis, transferred to PVDF and probed with anti-Histone H2A.Z (1:1000 dilution).
      Proteins were visualized using a rabbit anti-sheep secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).
      Biological Information
      ImmunogenLinear peptide corresponding the C-terminal amino acids 113-127 of human Histone H2A.Z.
      EpitopeC-terminus
      HostSheep
      SpecificityRecognizes Histone H2A.Z, Mr ~15 kDa.
      Species Reactivity
      • Human
      Species Reactivity NoteHuman. Wide species reactivity is expected.
      Predicted to cross-react with zebrafish, mouse, chicken, rat, rabbit, and Xenopus 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 encodes a member of the histone H2A family. Several transcript variants encoding different isoforms, have been identified for this gene. [provided by RefSeq].
      Gene Symbol
      • H2A.z
      • H2A/z
      • H2AZ
      • MGC117173
      • H2AFZ
      Purification MethodAffinity Purfied
      UniProt Number
      UniProt SummaryFUNCTION: Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. 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. May be involved in the formation of constitutive heterochromatin. May be required for chromosome segregation during cell division. Ref.7

      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. H2A or its variant H2AFZ forms an heterodimer with H2B. H2AFZ interacts with INCENP By similarity.

      SUBCELLULAR LOCATION:Nucleus.

      PTM: Monoubiquitination of Lys-122 gives a specific tag for epigenetic transcriptional repression.

      Acetylated on Lys-5, Lys-8 and Lys-12 during interphase. Acetylation disappears at mitosis By similarity.

      Not phosphorylated By similarity.

      SIMILARITY: Belongs to the histone H2A family.

      MOLECULAR WEIGHT: is 13413.4 Da from positions 2 - 128. Determined by ESI. Monoisotopic, not modified. Ref.8 Ref.10 Ref.11

      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 1 µg of either Normal Sheep IgG or 1 µg of Anti-Histone H2A.Z and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of H2A.Z associated DNA fragments was verified by qPCR using Control Primers (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: Variability 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 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDB

      Titel

      Sicherheitsdatenblatt (SDB) 

      Analysenzertifikate

      TitelChargennummer
      ChIPAb+ Histone H2A.Z2470777
      ChIPAb+ Histone H2A.Z - 21370352137035
      ChIPAb+ Histone H2A.Z - 20458632045863
      ChIPAb+ Histone H2A.Z - 22049472204947
      ChIPAb+ Histone H2A.Z - 23764692376469
      ChIPAb+ Histone H2A.Z - 24888242488824
      ChIPAb+ Histone H2A.Z - JBC1793871JBC1793871
      ChIPAb+ Histone H2A.Z - NG1827445NG1827445
      ChIPAb+ Histone H2A.Z - NRG1726316NRG1726316
      ChIPAb+ Histone H2A.Z -25640042564004

      Literatur

      ÜbersichtPub Med ID
      Altered nucleosome positioning at the transcription start site and deficient transcriptional initiation in Friedreich ataxia.
      Chutake, YK; Costello, WN; Lam, C; Bidichandani, SI
      The Journal of biological chemistry  289  15194-202  2014

      Abstract anzeigen
      24737321 24737321
      Distinctive patterns of epigenetic marks are associated with promoter regions of mouse LINE-1 and LTR retrotransposons.
      Rangasamy, D
      Mobile DNA  4  27  2013

      Abstract anzeigen
      24289137 24289137

      Broschüre

      Titel
      Advance your Epigenetics Research
      Product Selection Guide - Antibodies, small molecule inhibitors, kits, assays and proteins for signaling research.

      Technische Informationen

      Titel
      Guide to Chromatin Immunoprecipitation: Critical Factors for Success

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      Produktfamilien

      Kategorien

      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