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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, Vrt | WB, Cell Function Assay, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10121 |
| Trade Name |
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| Description | ChIPAb+ Acetyl-Histone H4 (Lys12) - ChIP Validated Antibody and Primer Set, rabbit monoclonal |
| Overview | All 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 (Lys12) set includes the Acetyl-Histone H4 (Lys12) antibody, a negative control normal rabbit IgG, and qPCR primers which amplify a 213 bp region of human GAPDH coding region. The Acetyl-Histone H4 (Lys12) 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 (Lys12) associated chromatin. |
| Alternate Names |
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| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Anti-Acetyl-Histone H4 (Lys12) (rabbit monoclonal), . One vial containing 50 µL of rabbit monoclonal IgG cell culture supernatant in 0.1% sodium azide. Store at -20°C. Negative Control Supernatant, . One vial containing 100 µL of cultured supernatant in 0.05% sodium azide. Store at -20°C. ChIP Primers, human GAPDH coding region,. One vial containing 75 μL of 5 μM each primer specific for human GAPDH coding region. Store at -20°C. FOR: GGC TCC CAC CTT TCT CAT CC REV: GGC CAT CCA CAG TCT TCT GG |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This ChIPAb+ Acetyl-Histone H4 (Lys12) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
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| 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: Representative lot data. 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) (Figure 3). |
| Biological Information | |
|---|---|
| Immunogen | Peptide corresponding to Histone H4 containing the sequence [GLG-AcK-GGA] on which Lys12 is acetylated. |
| Epitope | Unknown |
| Host | Rabbit |
| Specificity | This antibody detects histone H4 acetylated at Lys12. |
| Isotype | IgG |
| Species Reactivity |
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| Species Reactivity Note | Reacts with Human, wide range of cross-reactivity expected based on sequence homology. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | 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. |
| Gene Symbol |
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| Modifications |
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| Purification Method | Protein A Purfied |
| UniProt Number |
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| UniProt Summary | 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. |
| Molecular Weight | 11kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa S3 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 promoter (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 |
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| Storage and Shipping Information | |
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| Storage Conditions | Stable 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 Size | 25 assays |
| Material Package | 25 assays per set. Recommended use: 2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context). |