Histone H4 (K5,8,12ac) polyclonal antibody
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More Files
- More Functions
ChIP
ChIP assays were performed using human K562 cells. A titration of the antibody consisting of 0.2, 0.5, 1 and 2 ug per ChIP experiment was analysed. IgG (1 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. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ChIP-Seq
ChIP was performed on sheared chromatin from 100,000 K562 cells. The figure shows the signal distribution along the complete length of chromosome 2 and a zoomin to a 600 kb region and the enrichment in two genomic regions on chromosome 3 and 12, respectively, containing EIF4A2 and GAPDH positive controls.
Western Blot
Western Blot analysis of (1) 25 ug whole cell extracts of Hela cells, (2) 15 ug histone extracts of Hela cells, (3) 1 ug of recombinant histone H2A, (4) 1 ug of recombinant histone H2B, (5) 1 ug of recombinant histone H3, (6) 1 ug of recombinant histone H4.
Immunofluorescence
Immunofluorescent staining of Hela cell line with antibody followed by (A) an anti-rabbit antibody conjugated to Alexa488, (B) antibody after incubation of the antibody following blocking peptides: H4K5,8,12 unmodified, (C) H4K5,8,12ac, (D) H2A.ZK5,7,11ac, (E) H4K5,8,12,16ac (left). The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings (right).
Enzyme-linked Immunoabsorbent Assay
ELISA is a quantitative method used to determine the titer of the antibody using a serial dilution of antibody against Histone H4 (K5,8,12ac) 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 1:14500.
Dot Blot
Cross reactivity test using the Histone H4 (K5,8,12ac) antibody.
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 1:20000. The figure shows a high specificity of the antibody for the modification of interest.
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic peptide of Histone H4 (K5,8,12ac).
Immunogen
A synthetic peptide (conjugated with KLH) corresponding to Histone H4, acetylated at lysines 5, 8, and 12.
Host
Rabbit
Reactivity
Human, Mouse
Form
Liquid
Purification
Affinity purification
Recommend Usage
ELISA (1:1000)
Western Blot (1:1000)
ChIP (0.5-1 ug/IP)
Dot Blot (1:20000)
Immunofluorescence (1:500)
The optimal working dilution should be determined by the end user.Storage Buffer
In PBS (0.05% sodium azide, 0.05% proclin 300).
Storage Instruction
Store at -20°C. For long term storage store at -80°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
ChIP
ChIP assays were performed using human K562 cells. A titration of the antibody consisting of 0.2, 0.5, 1 and 2 ug per ChIP experiment was analysed. IgG (1 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. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).ChIP-Seq
ChIP was performed on sheared chromatin from 100,000 K562 cells. The figure shows the signal distribution along the complete length of chromosome 2 and a zoomin to a 600 kb region and the enrichment in two genomic regions on chromosome 3 and 12, respectively, containing EIF4A2 and GAPDH positive controls.Western Blot
Western Blot analysis of (1) 25 ug whole cell extracts of Hela cells, (2) 15 ug histone extracts of Hela cells, (3) 1 ug of recombinant histone H2A, (4) 1 ug of recombinant histone H2B, (5) 1 ug of recombinant histone H3, (6) 1 ug of recombinant histone H4.Immunofluorescence
Immunofluorescent staining of Hela cell line with antibody followed by (A) an anti-rabbit antibody conjugated to Alexa488, (B) antibody after incubation of the antibody following blocking peptides: H4K5,8,12 unmodified, (C) H4K5,8,12ac, (D) H2A.ZK5,7,11ac, (E) H4K5,8,12,16ac (left). The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings (right).Enzyme-linked Immunoabsorbent Assay
ELISA is a quantitative method used to determine the titer of the antibody using a serial dilution of antibody against Histone H4 (K5,8,12ac) 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 1:14500.Dot Blot
Cross reactivity test using the Histone H4 (K5,8,12ac) antibody.
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 1:20000. The figure shows a high specificity of the antibody for the modification of interest. -
Gene Info — HIST1H4A
Entrez GeneID
8359Protein Accession#
P62805Gene Name
HIST1H4A
Gene Alias
H4/a, H4FA
Gene Description
histone cluster 1, H4a
Omim ID
602822Gene Ontology
HyperlinkGene 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. [provided by RefSeq
Other Designations
H4 histone family, member A|histone 1, H4a
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