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ChIPAb+ Acetyl-Histone H4 (Lys5) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
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Species Reactivity Key Applications Host Format Antibody Type
H WB, IP, ChIP Rabbit Culture Supernatant Monoclonal Antibody
Description:
ChIPAb+ Acetyl-Histone H4 (Lys5) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
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Trade Name:
Upstate (Millipore)
Product 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.
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Specificity:
Recognizes Histone H4 acetylated on Lys5.
Molecular Weight:
~11 kDa
Epitope:
Acetylated Lys5
Immunogen:
Peptide corresponding to Histone H4 containing the sequence [GRG-AcK-GGK] on which Lys5 is acetylated.
Isotype:
IgG
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).
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Species Reactivity:
Human
Species Reactivity Note:
Human. Wide range of cross-reactivity expected based on sequence homology.
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 wi |