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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H | WB, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10047 |
| Brand Family | Upstate |
| Trade Name |
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| Description | ChIPAb+ Histone H4 - 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+ Histone H4 set includes the Histone H4 antibody, a negative control antibody (normal rabbit IgG), and qPCR primers which amplify a 110 bp region of human β-Globin. The Histone H4 and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H4-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. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Anti-Histone H4 (rabbit monoclonal IgG). One vial containing 50 µL of protein A purified rabbit monoclonal IgG supernatant in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide before addition of glycerol to 40%. Store at -20°C. Normal Rabbit IgG. One vial containing 75 µg of normal rabbit IgG in 75 µL storage buffer. Store at -20°C. ChIP Primers, β-Globin. One vial containing 75 μL of 5 μM of each primer specific for human β-Globin. Store at -20°C. FOR: AGG ACA GGT ACG GCT GTC ATC REV: TTT ATG CCC AGC CCT GGC TC |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This ChIPAb+ Histone H4 -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 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either normal rabbit IgG or 2 µL of Anti-Histone H4 antibody and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of total Histone H4 associated DNA fragments was verified by qPCR using ChIP Primers, β-Globin, as well as GAPDH promoter Control Primers, (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. HeLa Acid Extract (Lane 1 and 3), HeLa Acid Extract from sodium butyrate treated cells (Lane 2), HeLa Acid Extract from colcemid treated cells (Lane 4) and recombinant histone H4 (Lane 5) were resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-Histone H4 (1:1,000 dilution). Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and visualized using a chemiluminescence detection system. (Please see figures). |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated synthetic peptide corres-ponding to amino acids 17-28 of Histone H4. |
| Epitope | a.a 25-28 |
| Host | Rabbit |
| Specificity | Recognizes Histone H4. |
| Species Reactivity |
|
| Species Reactivity Note | 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. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H4 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in a histone cluster on chromosome 1. This gene is one of four histone genes in the cluster that are duplicated; this record represents the telomeric copy. [provided byRefSeq]. |
| Gene Symbol |
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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 | ~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 2 µL of either normal rabbit IgG or 2 µL of Anti-Histone H4 antibody and the Magna ChIP™ A Kit (Cat. #17-610). Successful immunoprecipitation of total Histone H4 associated DNA fragments was verified by qPCR using ChIP Primers, β-Globin (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. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage. |
| 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). |