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48-602MAG
Buffer Detection Kit for Magnetic Beads
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17-10045
Sigma-AldrichChIPAb+ Acetyl-Histone H4 (Lys5) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
This ChIPAb+ Acetyl-Histone H4 (Lys5) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
More>>This ChIPAb+ Acetyl-Histone H4 (Lys5) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. Less<<
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ChIPAb+ Acetyl-Histone H4 (Lys5) - 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 (Lys5) set includes the Acetyl-Histone H4 (Lys5) antibody, a negative control supernatant, and qPCR primers which amplify a 166 bp region of human GAPDH. The Acetyl-Histone H4 (Lys5) 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 (Lys5)-associated chromatin.
Alternate Names
H4K5Ac
Histone H4 (acetyl K5)
H4 histone family, member A
histone 1, H4a
histone cluster 1, H4a
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).
References
Product Information
Format
Culture Supernatant
Control
Includes negative control supernatant and primers specific for human GAPDH.
Presentation
Anti-Acetyl-Histone H4 (Lys5) (rabbit monoclonal IgG). One vial containing 50 μL of rabbit monoclonal IgG cell culture supernatant in 0.1% sodium azide. Store at -20°C.
Negative Control Supernatant (rabbit). One vial containing 50 μL of cultured supernatant in 0.05% sodium azide. Store at -20°C.
Control Primers. One vial containing 75 μL of 5 μM each primer specific for the human GAPDH promoter. Store at -20°C. FOR: TAC TAG CGG TTT TAC GGG CG REV: TCG AAC AGG AGG AGC AGA GAG CGA
This ChIPAb+ Acetyl-Histone H4 (Lys5) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
Western Blotting
Immunoprecipitation
Chromatin Immunoprecipitation (ChIP)
Application Notes
Chromatin Immunoprecipitation: Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either a negative control supernatant , or 2 µL of Anti-Acetyl-Histone H4 (Lys5) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of acetyl-Histone H4 (Lys5) associated DNA fragments was verified by qPCR using control Primers flanking the human GAPDH promoter, or primers amplifying the promoter of human β-globin, which is transcriptionally inactive in HeLa cells (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. Lysates from HeLa cells untreated or sodium butyrate treated (Lanes 1 and 2 respectively) were probed with anti-acetyl-Histone H4 (Lys5) (1:1,000 dilution). Proteins were visualized using goat anti-rabbit. (Please see figures).
Western Blot Analysis/Peptide Inhibition: Representative lot data. Sodium butyrate treated HeLa acid extract was resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-Acetyl-Histone H4 (Lys5) (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).
Biological Information
Immunogen
Peptide corresponding to Histone H4 containing the sequence [GRG-AcK-GGK] on which Lys5 is acetylated.
Epitope
Acetylated Lys5
Host
Rabbit
Specificity
Recognizes Histone H4 acetylated on Lys5.
Isotype
IgG
Species Reactivity
Human
Species Reactivity Note
Human. Wide range of cross-reactivity expected based on sequence homology.
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].
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
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Chromatin Immunoprecipitation: Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 µL of either negative control supernatant or 2 µL of Anti-Acetyl-Histone H4 (Lys5) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of acetyl-Histone H4 (Lys5) 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 ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
17-10045
04053252679247
Documentation
ChIPAb+ Acetyl-Histone H4 (Lys5) - ChIP Validated Antibody and Primer Set, rabbit monoclonal SDB
Millipore’s Histone H4 antibodies demonstrate specificity against linker histone H4. See below for H4 Antibodies, ChipAb+ Primer sets, and peptides, based on the expertise of Upstate & Chemicon. Weitere Informationen >>