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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, Yeast (S. cerevisiae) | WB, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10099 |
| Brand Family | Upstate |
| Trade Name |
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| Description | ChIPAb+ Acetyl-Histone H4 (Lys8) - 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 (Lys8) set includes the Acetyl-Histone H4 (Lys8) antibody, a negative control rabbit serum, and qPCR primers which amplify a 134 bp region of human HSPCA. The Acetyl-Histone H4 (Lys8) 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 (Lys8)-associated chromatin. |
| Alternate Names |
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| Background Information | Histone H4 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 H4 is involved with the structure of the nucleosomes of the beads on a string structure. The N-terminal tail of histone H4 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine. |
| Product Information | |
|---|---|
| Format | Serum |
| Control |
|
| Presentation | Anti-Acetyl-Histone H4 (Lys8) (rabbit polyclonal). One vial containing 50 μL of antiserum containing 0.05% sodium azide. Store at -20°C. Normal Rabbit Serum. One vial containing 100 μL of antiserum containing 0.05% sodium azide. Store at -20°C. ChIP Primers, HSPCA. One vial containing 75 μL of 5 μM of each primer specific for human HSPCA. Store at -20°C. FOR: GCA ACA GCT ACC ACA GGA CCA REV: GAG CGT GTG AAA TCA ACA TAA AGC |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | The ChIPAb+ Acetyl-Histone H4 (Lys8) set includes the Acetyl-Histone H4 (Lys8) antibody, a negative control rabbit serum & qPCR primers which amplify a 134 bp region of human HSPCA. |
| Key Applications |
|
| Application Notes | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 2 µL of Normal Rabbit Serum or 2 µL of Anti-Acetyl-Histone H4 (Lys8) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Acetyl-Histone H4 (Lys8) associated DNA fragments was verified by qPCR using ChIP Primers, human HSPCA for a positive, and known Negative Primers (Martinato, F., et al., 2008) as a negative assay (Please see figures). 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. Sodium Butyrate-treated HeLa acid extract was resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-Acetyl Histone H4 (Lys8) (1:1000 dilution). To demonstrate specificity, serum was preincubated with modified Histone H4 peptides: Lane 1: No peptide Lane 2: acetyl-Histone H4 (K5) (Cat.#:12-343) Lane 3: acetyl-Histone H4 (K8) (Cat.#:12-344) Lane 4: acetyl-Histone H4 (K12) (Cat.#:12-345) Lane 5: acetyl-Histone H4 (K16) (Cat.#:12-346) Lane 6: acetyl-Histone H4 (K5, 8, 12, 16) (Cat.#:12-353) Lane 7: Histone H4 (Cat.#:12-347) Proteins were visualized using a donkey anti-rabbit IgG conjugated to HRP and visualized using a chemiluminescence detection system (Please see figures). Immunoprecipitation: 5 μL of a previous lot immunoprecipitated acetyl Histone H4 from 1 mg of wild type whole cell yeast lysates but not from a yeast strain containing a Histone H4 Lys8R substitution. |
| Biological Information | |
|---|---|
| Immunogen | Synthetic peptide corresponding to yeast Histone H4 acetylated at lysine 8. |
| Host | Rabbit |
| Specificity | Recognizes acetyl-Histone H4 (Lys8). |
| Species Reactivity |
|
| Species Reactivity Note | Demonstrated to react with Yeast and human. Broad species cross-reactivity is expected based on 100% sequence homology. |
| Antibody Type | Polyclonal 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 | Unpurified |
| 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 | ~11 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 2 µL of Normal Rabbit Serum or 2 µL of Anti-Acetyl-Histone H4 (Lys8) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Acetyl-Histone H4 (Lys8) associated DNA fragments was verified by qPCR using ChIP Primers, human HSPCA (Please see figures). 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 | |
|---|---|
| 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). |