100 μL of rabbit monoclonal IgG cell culture supernatant with 0.1% sodium azide.
Applications
Application
This Anti-acetyl-Histone H4 (Lys12) Antibody, rabbit is validated for use in WB for the detection of acetyl-Histone H4 (Lys12).
Key Applications
Chromatin Immunoprecipitation (ChIP)
Western Blotting
Dot Blot
Application Notes
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 2 µL of Negative Control Supernatant, or 2 µL of Anti-Acetyl-Histone H4 (Lys12) and the Magna ChIP™ A Kit (Cat. # 17-610).
Successful immunoprecipitation of Acetyl-Histone H4 (Lys12)-associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH Coding Region as a positive locus, and a gene desert region 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:
Lysates from HeLa cells untreated or sodium butyrate treated (Lanes 1 and 2 respectively) were resolved probed with anti-acetyl-Histone H4 (Lys12) (1:1,000). Arrow indicates Acetyl-Histone H4 (Lys12).
Arrow indicates Acetyl-Histone H4 (Lys12) (~11 kDa)
Biological Information
Immunogen
Peptide corresponding to Histone H4 containing the sequence [GLG-AcK-GGA] on which Lys12 is acetylated.
Host
Rabbit
Specificity
Recognizes Histone H4 when acetylated on Lys12.
Isotype
IgG
Species Reactivity
Vertebrates Human
Species Reactivity Note
Reacts with Human, wide range of cross-reactivity expected based on sequence homology.
Histones 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 is intronless and encodes a member of the histone H4 family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6.
FUNCTION: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.
Product Usage Statements
Quality Assurance
Routinely evaluated by immunoblot.
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.
100 μL of rabbit monoclonal IgG cell culture supernatant with 0.1% sodium azide.
Applications
Application
This Anti-acetyl-Histone H4 (Lys12) Antibody, rabbit is validated for use in WB for the detection of acetyl-Histone H4 (Lys12).
Key Applications
Chromatin Immunoprecipitation (ChIP)
Western Blotting
Dot Blot
Application Notes
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 2 µL of Negative Control Supernatant, or 2 µL of Anti-Acetyl-Histone H4 (Lys12) and the Magna ChIP™ A Kit (Cat. # 17-610).
Successful immunoprecipitation of Acetyl-Histone H4 (Lys12)-associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH Coding Region as a positive locus, and a gene desert region 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:
Lysates from HeLa cells untreated or sodium butyrate treated (Lanes 1 and 2 respectively) were resolved probed with anti-acetyl-Histone H4 (Lys12) (1:1,000). Arrow indicates Acetyl-Histone H4 (Lys12).
Arrow indicates Acetyl-Histone H4 (Lys12) (~11 kDa)
Biological Information
Immunogen
Peptide corresponding to Histone H4 containing the sequence [GLG-AcK-GGA] on which Lys12 is acetylated.
Host
Rabbit
Specificity
Recognizes Histone H4 when acetylated on Lys12.
Isotype
IgG
Species Reactivity
Vertebrates Human
Species Reactivity Note
Reacts with Human, wide range of cross-reactivity expected based on sequence homology.
Histones 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 is intronless and encodes a member of the histone H4 family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6.
FUNCTION: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.
Product Usage Statements
Quality Assurance
Routinely evaluated by immunoblot.
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.
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