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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, Vrt, Yeast (S. cerevisiae) | WB, DB, ChIP, ChIP-seq | Rb | Serum | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 07-354 |
| Replaces | 04-1107 |
| Brand Family | Upstate |
| Trade Name |
|
| Description | Anti-acetyl-Histone H3 (Lys18) Antibody |
| Alternate Names |
|
| Background Information | Histone 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). |
| Product Information | |
|---|---|
| Format | Serum |
| Control |
|
| Presentation | 100 μLof rabbit serum containing 0.05% sodium azide and 30% glycerol. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Use Anti-acetyl-Histone H3 (Lys18) Antibody (Rabbit Polyclonal Antibody) validated in ChIP, WB to detect acetyl-Histone H3 (Lys18) also known as H3K18Ac, Histone H3 (acetyl K18). |
| Key Applications |
|
| Application Notes | Chromatin Immunoprecipitation: Representative lot data. Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either Normal Rabbit Serum , or 2 µL of Anti-Acetyl-Histone H3 (Lys18)and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Acetyl-Histone H3 (Lys18) associated DNA fragments was verified by qPCR using Control Primers specific for the human GAPDH promoter region as a positive locus, and β-globin primers as a negative locus. 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. ChIP-Sequencing: Representative lot data. Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 2 μg of anti-acetyl-Histone H3 (Lys18) antibody (cat# 07-354), 20 µL Protein A/g beads , and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq™ instrument. An excess of ten million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files. Western Blot Analysis: Representative lot data. Recombinant histone H3 (lane 1, Catalog # 14-494) and acid extracts from sodium butyrate treated (lane 2) and untreated (lane 3) HeLa cells (Catalog # 17-305) were probed with anti acetyl- Histone H3 (Lys18) (1:10,000 dilution). Arrow indicates acetyl histone H3 (Lys18) (17 kDa). DOT Blot: Representative lot data. 40 ng and 4ng amounts of histone peptides with various modifications (see table 1) were transferred to PVDF membrane and probed with Anti-Acetyl-Histone H3 (Lys18) antibody (1:5000 dilution). Proteins were visualized using a goat anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system. Image from a 60 second exposure is shown. |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated, synthetic peptide (GKAPRAcKQLASK-C) corresponding to amino acids 13-23 of yeast Histone H3 acetylated on lysine 18with a C-terminal cysteine added for conjugation purposes |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | Recognizes Histone H3 acetylated on lysine 18. |
| Species Reactivity |
|
| Species Reactivity Note | Yeast, Human, Most common vertebrates |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
|
| Entrez Gene Summary | Histones 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 H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. |
| Gene Symbol |
|
| Modifications |
|
| Purification Method | Unpurified |
| UniProt Number |
|
| UniProt Summary | FUNCTION: 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. TISSUE SPECIFICITY: Expressed in testicular cells. DEVELOPMENTAL STAGE: Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. 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 Weight | 17 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | routinely evaluated by immunoblot on acid extracts from sodium butyrate treated HeLa cells |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Maintain for 2 years at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |