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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, Eu | WB, Mplex, Cell Function Assay, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-630 |
| Brand Family | Upstate |
| Trade Name |
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| Description | ChIPAb+ Acetyl-Histone H4 - ChIP Validated Antibody and Primer Set |
| 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 set includes purified rabbit polyclonal antiserum and the normal rabbit IgG antiserum, which can be used to demonstrate that the acetyl-histone H4 antibody is capable of precipitating acetyl-histone H4 associated chromatin. The qPCR primers included flank the human GAPDH promoter and produce a 166 base pair PCR product. |
| Product Information | |
|---|---|
| Format | Serum |
| Control |
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| Presentation | Anti-Acetyl-Histone H4 (rabbit polyclonal IgG). One vial containing 100 μL of antiserum containing 0.05% sodium azide. Store at -20°C. Normal Rabbit Serum. One vial containing 100 uL antiserum containing 0.05% sodium azide. Store at -20°C. Control Primers. One vial containing 75 μL of 5 μM of each primer specific for for human GAPDH. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This ChIPAb+ Acetyl-Histone H4 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
|
| Application Notes | Western Blot Analysis: Acidextracted proteins from normal HeLa cells (Lane 1) and HeLa cells treated with 5 mM sodium butyrate for 24 hours (Lane 2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-acetyl Histone H4 (1:2000). Proteins were visualized using a goat-anti rabbit secondary antibody conjugated to HRP and a chemi-luminescence detection system. (Please see figures). |
| Biological Information | |
|---|---|
| Immunogen | The acetyl-histone H4 antiserum is made against a peptide corresponding to amino acids 2-19 of Tetrahymena histone H4. |
| Host | Rabbit |
| Specificity | Recognizes acetylated histone H4 of approximately 10 kDa. Cross-reacts with acetylated histone H2B from Tetrahymena and weakly crossreacts with acetylated histone H2B from HeLa cells. May cross-react with other acetylated proteins. |
| Species Reactivity |
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| Species Reactivity Note | Proven to react with human samples. Predicted broad cross-reactivity among eukaryotes based upon sequence similarity. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| UniProt Number |
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| UniProt Summary | Function: 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. Subunit structure: 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. Post-translational modification: Acetylation at Lys-6 (H4K5ac), Lys-9 (H4K8ac), Lys-13 (H4K12ac) and Lys-17 (H4K16ac) occurs in coding regions of the genome but not in heterochromatin. Citrullination at Arg-4 (H4R3ci) by PADI4 impairs methylation. Monomethylation at Arg-4 (H4R3me1) by PRMT1 favors acetylation at Lys-9 (H4K8ac) and Lys-13 (H4K12ac). Demethylation is performed by JMJD6. Symmetric dimethylation on Arg-4 (H4R3me2a) by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage. Monomethylated, dimethylated or trimethylated at Lys-21 (H4K20me1, H4K20me2, H4K20me3). Monomethylation is performed by SET8. Trimethylation is performed by SUV420H1 and SUV420H2 and induces gene silencing. 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. Monoubiquitinated at Lys-92 of histone H4 (H4K91ub1) in response to DNA damage. The exact role of H4K91ub1 in DNA damage response is still unclear but it may function as a licensing signal for additional histone H4 post-translational modifications such as H4 Lys-21 methylation (H4K20me). Sumoylated, which is associated with transcriptional repression. Sequence similarities: Belongs to the histone H4 family. |
| Molecular Weight | 10 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents) was subjected to chromatin immunoprecipitation using 4 μL of either a normal rabbit antiserum or Anti-Acetyl Histone H4 serum and the Magna ChIP A (Cat. # 17-610) Kit. Successful immunoprecipitation of acetyl histone H4 associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter (Please see figures). Please refer to the EZ-Magna A ChIP™ (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -20°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 25 assays |
| Material Package | 25 assays per set. ~4 μL per chromatin immunoprecipitation |