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17-10045 ChIPAb+ Acetyl-Histone H4 (Lys5) - ChIP Validated Antibody and Primer Set, rabbit monoclonal

This ChIPAb+ Acetyl-Histone H4 (Lys5) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Applications: Western Blotting, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP) Research Category: Epigenetics & Nuclear Function Research Sub-Category: Histones
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17-10045
25 assays  25 assays per set. Recommended use: ~2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

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      Key Spec Table

      Species ReactivityKey Applications
      HWB, IP, ChIP
      Description
      Catalogue Number17-10045
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ Acetyl-Histone H4 (Lys5) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
      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+ Acetyl-Histone H4 (Lys5) set includes the Acetyl-Histone H4 (Lys5) antibody, a negative control supernatant, and qPCR primers which amplify a 166 bp region of human GAPDH. The Acetyl-Histone H4 (Lys5) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Acetyl-Histone H4 (Lys5)-associated chromatin.
      Alternate Names
      • H4K5Ac
      • Histone H4 (acetyl K5)
      • H4 histone family, member A
      • histone 1, H4a
      • histone cluster 1, H4a
      Background InformationHistone H4 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 H4 is involved with the structure of the nucleosomes of the 'beads on a string' structure. Acetylation of histone H4 occurs at several different lysine positions in the histone tail and is performed by a family of enzymes known as Histone Acetyl Transferases (HATs).
      Product Information
      FormatCulture Supernatant
      Control
      • Includes negative control supernatant and primers specific for human GAPDH.
      PresentationAnti-Acetyl-Histone H4 (Lys5) (rabbit monoclonal IgG). One vial containing 50 μL of rabbit monoclonal IgG cell culture supernatant in 0.1% sodium azide. Store at -20°C.

      Negative Control Supernatant (rabbit). One vial containing 50 μL of cultured supernatant in 0.05% sodium azide. Store at -20°C.

      Control Primers. One vial containing 75 μL of 5 μM each primer specific for the human GAPDH promoter. Store at -20°C.
      FOR: TAC TAG CGG TTT TAC GGG CG
      REV: TCG AAC AGG AGG AGC AGA GAG CGA
      Quality LevelMQ100
      Applications
      ApplicationThis ChIPAb+ Acetyl-Histone H4 (Lys5) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Western Blotting
      • Immunoprecipitation
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either a negative control supernatant , or 2 µL of Anti-Acetyl-Histone H4 (Lys5) and the Magna ChIP™ A Kit (Cat. # 17-610).
      Successful immunoprecipitation of acetyl-Histone H4 (Lys5) associated DNA fragments was verified by qPCR using control Primers flanking the human GAPDH promoter, or primers amplifying the promoter of human β-globin, which is transcriptionally inactive in HeLa cells (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.
      Lysates from HeLa cells untreated or sodium butyrate treated (Lanes 1 and 2 respectively) were probed with anti-acetyl-Histone H4 (Lys5) (1:1,000 dilution).
      Proteins were visualized using goat anti-rabbit. (Please see figures).

      Western Blot Analysis/Peptide Inhibition: Representative lot data.
      Sodium butyrate treated HeLa acid extract was resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-Acetyl-Histone H4 (Lys5) (1:1000 dilution).
      To demonstrate specificity, serum was preincubated with modified Histone H4 peptides:

      Lane 1: No peptide
      Lane 2: Acetyl-Histone H4 (K5) (Cat. # 12-343)
      Lane 3: Acetyl-Histone H4 (K8) (Cat. # 12-344)
      Lane 4: Acetyl-Histone H4 (K12) (Cat. # 12-345)
      Lane 5: Acetyl-Histone H4 (K16) (Cat. # 12-346)
      Lane 6: Acetyl-Histone H4 (K5, 8, 12, 16) (Cat. # 12-353)
      Lane 7: Histone H4 (Cat. # 12-347)

      Proteins were visualized using a donkey anti-rabbit IgG conjugated to HRP and visualized using a chemiluminescence detection system. (Please see figures).
      Biological Information
      ImmunogenPeptide corresponding to Histone H4 containing the sequence [GRG-AcK-GGK] on which Lys5 is acetylated.
      EpitopeAcetylated Lys5
      HostRabbit
      SpecificityRecognizes Histone H4 acetylated on Lys5.
      IsotypeIgG
      Species Reactivity
      • Human
      Species Reactivity NoteHuman. Wide range of cross-reactivity expected based on sequence homology.
      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. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an 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 H4 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in a histone cluster on chromosome 1. This gene is one of four histone genes in the cluster that are duplicated; this record represents the telomeric copy. [provided byRefSeq].
      Gene Symbol
      • HIST1H4A
      • H4/J
      • HIST1H4F
      • HIST1H4L
      • H4FN
      • H4FA
      • H4FH
      • HIST1H4H
      • H4F2
      • H4FM
      • H4/A
      • H4FK
      • H4FG
      • HIST2H4
      • H4FB
      • H4/K
      • HIST1H4I
      • H4/N
      • HIST1H4B
      • H4FE
      • H4/M
      • H4/a
      • HIST1H4E
      • HIST4H4
      • HIST2H4A
      • H4FC
      • H4FI
      • H4/H
      • HIST1H4C
      • H4/G
      • HIST1H4K
      • H4FJ
      • H4FD
      • H4/I
      • H4/B
      • H4/D
      • H4/C
      • HIST1H4J
      • HIST1H4D
      • H4/E
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P62805 # 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: 103 amino acids; 11367 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: Symmetric dimethylation on Arg-4 by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage (By similarity). & Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins.
      SIMILARITY: SwissProt: P62805 ## Belongs to the histone H4 family.
      Molecular Weight~11 kDa
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 µL of either negative control supernatant or 2 µL of Anti-Acetyl-Histone H4 (Lys5) and the Magna ChIP™ A Kit (Cat. # 17-610).
      Successful immunoprecipitation of acetyl-Histone H4 (Lys5) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter (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.
      Packaging Information
      Material Size25 assays
      Material Package25 assays per set. Recommended use: ~2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Global Trade Item Number
      Catalogue Number GTIN
      17-10045 04053252679247