Histone H3 (K9/14ac) polyclonal antibody

Catalog # PAB31269

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Size:50 ug
Price: USD $ 638.00
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Contact Info
  • +1-909-264-1399
    +1-909-992-0619
    Toll Free : +1-877-853-6098
  • +1-909-992-3401
Images
ChIP
Application

ChIP

ChIP assays were performed using human HeLa cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (2 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as positive controls, and for the coding region of the inactive MB gene and the Sat2 satellite repeat, 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
Application

ChIP-Seq

ChIP was performed on sheared chromatin from 100,000 K562 cells using 1 ug of antibody. IgG (1 ug/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the promoters of the active GAPDH and EIF4A2 genes, used as positive control targets, and the coding region of the inactive MB gene and the Sat2 satellite repeat, used as negative control targets. The figure shows the peak distribution along the complete sequence and a 1.5 Mb region of the X-chromosome and in two regions surrounding the GAPDH and EIF4A2 positive control genes. The position of the amplicon used for ChIP-qPCR is indicated by an arrow. These results clearly show an enrichment of the H3K9/14 acetylation at the promoters of active genes.

Western Blot (Cell lysate)
Application

Western Blot (Cell lysate)

Western Blot (Cell lysate) 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, and (6) 1 ug of recombinant histone H4.

Immunofluorescence
Application

Immunofluorescence

Immunofluorescent staining of Hela cell line with antibody followed by an anti-rabbit antibody conjugated to Alexa488 (left). The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings (right).

Enzyme-linked Immunoabsorbent Assay
Application

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 H3 (K9/14ac). 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 1:4000.

Dot Blot
Application

Dot Blot

Cross reactivity tests using the Histone H3 (K9/14ac) antibody.
Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K9. 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.

  • Specification

    Product Description

    Rabbit polyclonal antibody raised against synthetic peptide of Histone H3 (K9/14ac).

    Immunogen

    A synthetic peptide (conjugated with KLH) corresponding to Histone H3, acetylated lysines at 9 and 14.

    Host

    Rabbit

    Reactivity

    Human, Mouse, Rainbow trout, Zebra fish, A. Nidulans, Arabidopsis

    Form

    Liquid

    Purification

    Affinity purification

    Recommend Usage

    ELISA (1:100)
    Western Blot (1:1000)
    ChIP (1-2 ug/CHIP)
    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.

  • Applications

    ChIP

    ChIP assays were performed using human HeLa cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (2 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as positive controls, and for the coding region of the inactive MB gene and the Sat2 satellite repeat, 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 using 1 ug of antibody. IgG (1 ug/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the promoters of the active GAPDH and EIF4A2 genes, used as positive control targets, and the coding region of the inactive MB gene and the Sat2 satellite repeat, used as negative control targets. The figure shows the peak distribution along the complete sequence and a 1.5 Mb region of the X-chromosome and in two regions surrounding the GAPDH and EIF4A2 positive control genes. The position of the amplicon used for ChIP-qPCR is indicated by an arrow. These results clearly show an enrichment of the H3K9/14 acetylation at the promoters of active genes.

    Western Blot (Cell lysate)

    Western Blot (Cell lysate) 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, and (6) 1 ug of recombinant histone H4.

    Immunofluorescence

    Immunofluorescent staining of Hela cell line with antibody followed by an anti-rabbit antibody conjugated to Alexa488 (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 H3 (K9/14ac). 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 1:4000.

    Dot Blot

    Cross reactivity tests using the Histone H3 (K9/14ac) antibody.
    Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K9. 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 — HIST1H3A

    Entrez GeneID

    8350

    Protein Accession#

    P68431

    Gene Name

    HIST1H3A

    Gene Alias

    H3/A, H3FA

    Gene Description

    histone cluster 1, H3a

    Omim ID

    602810

    Gene Ontology

    Hyperlink

    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 H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq

    Other Designations

    H3 histone family, member A|histone 1, H3a

  • Interactome
  • Pathway
  • Publication Reference
Contact Info
  • +1-909-264-1399
    +1-909-992-0619
    Toll Free : +1-877-853-6098
  • +1-909-992-3401
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