Millipore Sigma Vibrant Logo
Attention: We have moved. EMD Millipore products are no longer available for purchase on emdmillipore.com.Learn More

17-10241 ChIPAb+™ Acetyl-Histone H3 (Lys9/18) - ChIP Validated Antibody and Primer Set

View Products on Sigmaaldrich.com
17-10241
25 assays  25 assays per set. Recommended use: ~1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
Purchase on Sigma-Aldrich

Special Offers

Overview

Replacement Information

Key Specifications Table

Species ReactivityKey Applications
B, H, M, Yeast (S. cerevisiae)WB, ChIP
Description
Catalogue Number17-10241
Trade Name
  • ChIPAb+
  • Upstate
DescriptionChIPAb+™ Acetyl-Histone H3 (Lys9/18) - 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+ Acetyl-Histone H3 (Lys9/18) set includes the Acetyl-Histone H3 (Lys9/18) antibody, a Normal rabbit serum, and control primers which amplify a 166 bp region of human GAPDH promoter. The Acetyl-Histone H3 (Lys9/18) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Acetyl-Histone H3 (Lys9/18)-associated chromatin.
Alternate Names
  • H3K9/18Ac
  • Histone H3 (acetyl K9/18)
Background InformationHistone H3 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 H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure.

Acetylation of histone H3 occurs at several different lysine positions in the histone tail and is performed by a family of enzymes known as Histone Acetyl Transferases (HATs).
References
Product Information
FormatSerum
Control
  • Includes normal rabbit serum and primers specific for human GAPDH promoter.
PresentationAnti-Acetyl-Histone H3 (Lys9/18) (rabbit polyclonal), . One vial containing 25 µL of rabbit antiserum containing 0.05% 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.
Control Primers, human GAPDH promoter, . 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
Quality LevelMQ100
Applications
ApplicationThis ChIPAb+ Acetyl-Histone H3 (Lys9/18) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
  • Western Blotting
  • 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-Acetyl-Histone H3 (Lys9/18), and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of acetyl-Histone H3 (Lys9/18) associated DNA fragments was verified by qPCR using Control Primers specific for the human GAPDH promoter region as a positive locus and human β-globin primers 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 extracts from untreated (lane 1), sodium butyrate treated (lane 2, Catalog # 17-305) HeLa cells and recombinant Histone
H3, (Lane 3, Catalog # 14-411) were resolved by electrophoresis, transferred to nitrocellulose and probed with Anti-Acetyl-Histone H3 (Lys9/18) (1:5,000 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates Acetyl-histone H3 (Lys9/18) (~17kDa). (Figure 3).
Beadlyte® Histone-Peptide Specificity Assay:
Representative lot data.
1:1,000-1:40,000 dilutions of anti-acetyl-Histone H3 (Lys9/18) were incubated with a cocktail of microspheres conjugated to histone H3 peptides with the following modifications:
1. acetyl-lysine 4
2. acetyl-lysine 14
3. acetyl-lysine 9
4. acetyl-lysine 18
5. No modifications, containing aa 1-20
Unbound antibody was removed by filtration. The peptide-antibody complexes were incubated with a biotin-conjugated anti-mouse secondary antibody followed by incubation with a phycoerythrin-streptavidin conjugate.
Fluorescence was read on a Luminex® 100™ instrument. Median
fluorescence intensity (MFI) is plotted. (Figure 4).

Biological Information
ImmunogenKLH-conjugated synthetic peptide (..ARAcKSTGGKAPRAcKQL..) in which AcK corresponds to acetyl-lysine at residue 9 and 18 of human Histone H3.
EpitopeAcetylated Lys9/18
HostRabbit
SpecificityRecognizes Histone H3 acetylated on lysines 9 and 18.
Species Reactivity
  • Bovine
  • Human
  • Mouse
  • Yeast (S. cerevisiae)
Species Reactivity NoteBroad species cross-reactivity expected
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
  • H3F3A
  • MGC87783
  • H3.3A
  • MGC87782
  • H3F3
  • H3.3B
  • H3F3B
Modifications
  • Acetylation
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 Weight17 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
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-Acetyl-Histone H3 (Lys9/18) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Acetyl-Histone H3 (Lys9/18) associated DNA fragments was verified by qPCR using Control Primers specific for the human GAPDH promoter region as a positive locus (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).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalog Number GTIN
17-10241 04053252556166

Documentation

ChIPAb+™ Acetyl-Histone H3 (Lys9/18) - ChIP Validated Antibody and Primer Set Certificates of Analysis

TitleLot Number
ChIPAb+ Acetyl-Histone H3 (Lys9/18) - 1992289 1992289
ChIPAb+ Acetyl-Histone H3 (Lys9/18) - 2053034 2053034
ChIPAb+ Acetyl-Histone H3 (Lys9/18) - NRG1880394 NRG1880394
ChIPAb+™ Acetyl-Histone H3 (Lys9/18) 3171005
ChIPAb+™ Acetyl-Histone H3 (Lys9/18) Polyclonal Antibody/Primer Set 3766912
ChIPAb+™ Acetyl-Histone H3 (Lys9/18) Polyclonal Antibody/Primer Set 2869079
ChIPAb+™ Acetyl-Histone H3 (Lys9/18) Polyclonal Antibody/Primer Set 2965346
ChIPAb+™ Acetyl-Histone H3, (Lys9/18), Polyclonal Antibody/Primer Set 2837839

References

Reference overviewPub Med ID
An ARID domain-containing protein within nuclear bodies is required for sperm cell formation in Arabidopsis thaliana.
Zheng, B; He, H; Zheng, Y; Wu, W; McCormick, S
PLoS genetics  10  e1004421  2014

Show Abstract
25057814 25057814

Brochure

Title
Advance your Epigenetics Research

Related Products & Applications

Product Families

Related Products By: Brand Facete

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