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Synthetic peptide within Human Histone H2A (acetyl K5) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
Our Abpromise guarantee covers the use of ab195486 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|CHIPseq||Use at an assay dependent concentration.
Use 0.5 μg per CHIPseq reaction.
|WB||1/1000. Predicted molecular weight: 14 kDa.|
|ChIP||Use at an assay dependent concentration.
Use 0.5-5 µg per ChIP
ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using 0.5 μg of ab195486. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. This figure shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (panels A and B) and in genomic regions of chromosome 7, surrounding the ACTB gene, and of chromosome 12, surrounding the GAPDH gene (panels C and D). The position of the amplicon used for ChIP-qPCR is indicated by an arrow.
Immunofluorescent analysis of HeLa cells (4% formaldehyde-fixed) labeling Histone H2A (acetyl K5) with ab195486 at 1/500 dilution followed by an anti-rabbit antibody conjugated to Alexa488 (left panel). The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
Dot Blot analysis was performed to test the cross reactivity of ab195486 against Histone H2A (acetyl K5) with peptides containing other histone modifications and the unmodified H2AK5. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. ab195486 was used at 1/5,000 dilution.
ChIP analysis using HeLa cells, ab195486 and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH and ACTB promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls.
ab195486 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"