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17-10254 ChIPAb+™ Histone H3 (CT) - ChIP Validated Antibody and Primer Set

This ChIPAb+ Histone H3 (C-Term) -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, Immunocytochemistry, Dot Blot, Peptide Inhibition Assay, Chromatin Immunoprecipitation (ChIP) Research Category: Epigenetics & Nuclear Function Research Sub-Category: Histones
17-10254
25 assays  25 assays per set. Recommended use: 1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
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Key Spec Table

Species ReactivityKey Applications
Ch, H, M, R, Yeast (S. cerevisiae)WB, ICC, DB, PIA, ChIP
Description
Catalogue Number17-10254
Trade Name
  • ChIPAb+
  • Upstate
DescriptionChIPAb+™ Histone H3 (CT) - 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 H3 (C-Term) set includes the Histone H3 (C-Term) antibody, a Normal Rabbit Serum, and control primers which amplify a 110 bp region of ChIP Primers, human β-globin. The Histone H3 (C-Term) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H3 (C-Term)-associated chromatin.
Alternate Names
  • Histone H3.1t
  • H3t
  • histone 3
  • H3/g
Background InformationHistone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.
Product Information
FormatSerum
Control
  • Includes normal rabbit serum and primers specific for human β-globin.
PresentationAnti-Histone H3 (C-Term) (rabbit polyclonal). One vial containing 25 µL of rabbit antiserum diluted 1:2 in storage buffer (0.02 M Phosphate, 0.25 M NaCl, 0.1% sodium azide) before the addition of glycerol to 30%. Store at -20°C.
Normal Rabbit Serum. One vial containing 25 μL of antiserum containing 0.05% sodium azide. Store at -20°C.
ChIP Primers, human 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
ApplicationThis ChIPAb+ Histone H3 (C-Term) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
  • Western Blotting
  • Immunocytochemistry
  • Dot Blot
  • Peptide Inhibition Assay
  • Chromatin Immunoprecipitation (ChIP)
Application NotesChromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Rabbit Serum,or 1 µL of Anti-Histone H3 (C-Term) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of histone H3 associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin as a positive locus, and GAPDH promoter primers (22-004) as a negative locus. (Figure 2). 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.
Acid extracted proteins from HeLa cells untreated (lane 1) or treated with sodium butyrate (lane 2) or colcemid (lane 3) and recombinant Histone H3 (lane 4) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Histone H3, CT, pan (1:50,000 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates Histone H3 (~17 kDa). (Figure 3).
Biological Information
ImmunogenKLH-conjugated, synthetic peptide corresponding to the C-terminus of human Histone H3.
EpitopeC-terminus
HostRabbit
SpecificityRecognizes C-Terminal region of Histone H3, Mr 17 kDa
Species Reactivity
  • Chicken
  • Human
  • Mouse
  • Rat
  • Yeast (S. cerevisiae)
Species Reactivity NoteHuman, mouse, rat, yeast and chicken. Broad species cross-reactivity expected due to 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. 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 contains introns and its mRNA is polyadenylated, unlike most histone genes. The protein encoded is a replication-independent member of the histone H3 family.
Gene Symbol
  • HIST3H3
  • H3FT
  • MGC126886
  • H3t
  • MGC126888
  • H3T
  • H3/g
  • H3.4
  • H3/t
Purification MethodUnpurified
UniProt Number
UniProt SummaryFUNCTION: SwissProt: Q16695 # 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: 136 amino acids; 15508 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: Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18 (By similarity). & Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription (By similarity). & Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression (By similarity). & Methylation at Lys-5, Lys-37 and Lys-80 are linked to gene activation. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28 are linked to gene repression. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin (By similarity). & Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase. Phosphorylated at Ser-11 during the whole mitosis. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation (By similarity). & Phosphorylation at 'Ser-11' is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' is important during interphase because it enables the transcription of genes following external stimulation, like stress or growth factors. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylation at 'Ser-11' by AURKB/Aurora-B mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. & Ubiquitinated (By similarity).
SIMILARITY: SwissProt: Q16695 ## Belongs to the histone H3 family.
Molecular WeightApprox. 17 kDa
Product Usage Statements
Quality AssuranceChromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Rabbit Serum, or 1 µL of Anti-Histone H3 (C-Term) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of histone H3 associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin (Figure 1).
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 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
Global Trade Item Number
Catalogue Number GTIN
17-10254 04053252592355