Anti-Histone H3 (acetyl K4) antibody (ab232931)
Key features and details
- Rabbit polyclonal to Histone H3 (acetyl K4)
- Suitable for: ChIP, ChIP-sequencing, WB, ICC/IF, Dot blot
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Histone H3 (acetyl K4) antibody
See all Histone H3 primary antibodies -
Description
Rabbit polyclonal to Histone H3 (acetyl K4) -
Host species
Rabbit -
Tested applications
Suitable for: ChIP, ChIP-sequencing, WB, ICC/IF, Dot blotmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human Histone H3 (acetyl K4) conjugated to keyhole limpet haemocyanin.
Database link: P68431 -
Positive control
- ChIP: Chromatin from HeLa cells. ChIPseq: Chromatin from HeLa cells. WB: HeLa whole cell and histone extracts. ICC/IF: HeLa cells.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservatives: 0.05% Sodium azide, 0.05% Proclin 300
Constituent: PBS -
Concentration information loading...
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Purity
Affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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ChIP was performed with 1 μg ab232931 against Histone H3 (acetyl K4) on sheared chromatin from 4 million HeLa (human epithelial cell line from cervix adenocarcinoma) cells. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. The figure shows the peak distribution along the complete sequence and a 1.5 mb region of the X chromosome (A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes (C and D).
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ChIP assays were performed using HeLa (human epithelial cell line from cervix adenocarcinoma) cells, ab232931 against Histone H3 (acetyl K4) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 4 million cells. A titration of ab232931 consisting of 1, 2, 5 and 10 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter 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).
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Dot Blot analysis was performed to test the cross reactivity of ab232931 against Histone H3 (acetyl K4) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. ab232931 was used at 1/5000 dilution. ab232931 shows a high specificity for the modification of interest.
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All lanes : Anti-Histone H3 (acetyl K4) antibody (ab232931) at 1/1000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell extract at 25 µg
Lane 2 : HeLa histone extract at 15 µg
Predicted band size: 15 kDa
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HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for Histone H3 (acetyl K4) (green) using ab232931 at 1/200 dilution in ICC/IF. 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 ab232931 (left) at 1/500 dilution 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.