Histone H4 Antibody (Acetyl-Lys5/8/12/16) (OADC00138)

Data Sheet
 
Product Number OADC00138
Product Page www.avivasysbio.com/histone-h4-antibody-acetyl-lys5-8-12-16-oadc00138.html
Name Histone H4 Antibody (Acetyl-Lys5/8/12/16) (OADC00138)
Conjugation Unconjugated
NCBI Gene Id 121504; 554313; 8294; 8359; 8360; 8361; 8362; 8363; 8364; 8365; 8366; 8367; 8368; 8370
Host Rabbit
Clonality Polyclonal
Concentration 0.76 mg/mL
Gene Full Name histone cluster 4, H4
Alias Symbols H4/p, H4C1, H4C2, H4C3, H4C4, H4C5, H4C6, H4C8, H4C9, H4C11, H4C12, H4C13, H4C14, H4C15, HIST4H4
Product Format Liquid. Provided in PBS containing 0.05% azide and 0.05% ProClin 300.
Description of Target Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone 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 replication-dependent histone that is a member of the histone H4 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element.
Reconstitution and Storage Store at -20C. Avoid repeated freeze/thaw cycles.
Datasheets/Manuals Printable datasheet for anti-HIST4H4 (OADC00138) antibody
Specificity Polyclonal antibody raised in rabbit against the region of histone H4 containing the acetylated lysines 5, 8, 12 and 16 (H4K5,8,12,16ac), using a KLH-conjugated synthetic peptide.
Application Info CHIP: 2 ug per IP
ELISA: 1:1,000
Dot Blotting: 1:20,000
IF: 1:500
Uniprot ID P62805
Protein Name histone H4
Protein Accession # NP_001029249.1
Purification Affinity purified
Nucleotide Accession # NM_001034077.4
Target Post-Translational Modification Acetyl-Lys5/8/12/16
Preservative 0.05% Sodium Azide and 0.05% ProClin 300.
Gene Symbol HIST4H4
Predicted Species Reactivity Human
Application CHIP, DB, IF
Predicted Homology Based on Immunogen Sequence Yeast
Image 1
Human Hela cells
ChIP results obtained with the Aviva antibody directed against H4K5,8,12,16ac. ChIP assays were performed using human HeLa cells, the Aviva antibody against H4K5,8,12,16ac and optimized PCR primer sets for qPCR. ChIP was performed with the "iDeal ChIP- seq" kit on sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 ug per ChIP experiment was analysed. IgG (2 ug/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active gene EIF4A2 and for a region 1 kb upstream of the GAPDH gene, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Image 2
Human Hela cells
ChIP-seq results obtained with the Aviva antibody directed against H4K5,8,12,16ac r. ChIP was performed with 2 ug of the Aviva antibody against H4K5,8,12,16ac on sheared chromatin from 1 million HeLa cells. The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure shows the signal distribution along the complete length of chromosome 5 and a zoomin to a 600 kb region. Figures show the enrichment in two genomic regions on chromosome 3 and 12, respectively, containing EIF4A2 and GAPDH positive controls. The position of the amplicon used for validating the QPCR results is shown with an arrow
Image 3
H4K5,8,12,16ac
Cross reactivity tests using the Aviva antibody directed against H4K5,8,12,16ac. To test the cross reactivity of the Aviva antibody against H4K5,8,12,16ac, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 4 shows a high specificity of the antibody for the modification of interest.
Image 4
H4K5,8,12,16ac
Determination of the antibody titer. To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Aviva antibody directed against H4K5,8,12,16ac in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:21,200.
Image 5
Mouse NIH3T3 cells
Immunofluorescence using the Aviva antibody directed against H4K5,8,12,16ac. Mouse NIH3T3 cells were stained with the Aviva antibody against H4K5,8,12,16ac and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K5,8,12,16ac antibody (left) diluted 1:500 in blocking solution followed by an anti- rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
 

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