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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, Vrt | IP, WB, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10101 |
| Brand Family | Upstate |
| Trade Name |
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| Description | ChIPAb+ Acetyl-Histone H4 (Lys16) - 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 (Lys16) set includes the Acetyl-Histone H4 (Lys16) antibody, a negative control normal rabbit IgG, and qPCR primers which amplify a 178 bp region of human RPL10 promoter. The Acetyl-Histone H4 (Lys16) 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 (Lys16)-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. Acetylation of histone H4 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 | Purified |
| Control |
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| Presentation | Anti-Acetyl-Histone H4 (Lys16) (rabbit polyclonal). One vial containing 50 µL of affinity purified rabbit polyclonal serum in buffer containing 0.1 M Tris-Glycine (pH 7.4) 150 mM NaCl with 0.05% sodium azide. Store at -20°C. Normal Rabbit IgG. One vial containing 125 µg of rabbit IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C. ChIP Primers, RPL10 promoter. One vial containing 75 μL of 5 μM of each primer specific for the promoter region of human RPL10. Store at -20°C. FOR: ACC CGT CTT CGA CAG GAC T REV: GGA ACG GAA GAC GAG AAC AG |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This ChIPAb+ Acetyl-Histone H4 (Lys16) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
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| 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 either 2 µg of Normal rabbit IgG or 2 µL of Anti-acetyl-Histone H4 (Lys16) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of acetyl-Histone H4 (Lys16) associated DNA fragments was verified by qPCR using ChIP Primers, RPL10 Promoter as a positive locus, and β-Globin primers as a negative locus. (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. Recombinant Histone H4 (Cat # 14-697) (lane1) and HeLa acid extract (lane 2) was resolved by electrophoresis, transferred to PVDF and probed with anti-Acetyl Histone H4 (Lys16) (0.1 μg/mL). Proteins were visualized using a donkey anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Acetyl Histone H4 (Lys16) (~10 kDa) (Please see figures). Multiplexing (Luminex®): Representative lot data. This antibody specifically recognizes histone H4 acetylated on Lys16 by Luminex assay (Please see figures). Peptide Inhibition Assay: Representative lot data. This antibody peptide blocked on HeLa cell extracts (Please see figures). |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated Histone H4 corresponding to the N-terminus region at and around acetylated Lys16. |
| Epitope | Acetylated H4 Lys16 |
| Host | Rabbit |
| Specificity | This antibody detects histone H4 acetylated at Lys16. |
| Species Reactivity |
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| Species Reactivity Note | Human. Broad species cross-reactivity is expected based on 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 | Affinity 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 | ~10 kDa observed |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 2 µg of Normal Rabbit IgG or 2 µL of Anti-acetyl-Histone H4 (Lys16) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of acetyl-Histone H4 (Lys16) associated DNA fragments was verified by qPCR using ChIP Primers, RPL10 Promoter (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). |