ChIP for low abundance proteins: ChIP-IT High Sensitivity

The ChIP-IT High Sensitivity® (HS) kit from Active Motif is the most sensitive ChIP kit available on the market. Designed and optimised to provide superior results when performing ChIP using antibodies against low abundance proteins, such as transcription factors, the ChIP-IT HS kit also enables high quality DNA enrichment from as little as 1000 cells per ChIP reaction for abundant targets.

Benefits of using the ChIP-IT HS kit 
• Ideal for low abundance transcription factors or antibodies with low binding affinities
• Sensitive enrichment of DNA from >1000 cells/reaction for high abundance target proteins
• Sensitive enrichment of DNA from >50000 cells/reaction for low abundance transcription factors
• Optimised reagents reduce background levels caused by non-specific binding events
• Filtration based washes offer a faster, easier solution with better consistency than magnetic capture
• Highly robust procedure validated across multiple sample types 
• Proven performance in both qPCR and ChIP-Seq analysis

Detect low abundance protein targets with ChIP-IT HS
The ChIP-IT HS kit is ideal for use with challenging antibodies that do not give signal with other ChIP methods, as the method is sensitive enough to detect specific binding of even low abundance proteins. Below is a comparison of ChIP-IT High Sensitivity with other commercially available ChIP kits.

Comparison of ChIP kits targeting a low abundance transcription factor. MCF-7 chromatin was prepared according to each manufacturer's recommendations for their ChIP assay from 1.5 x 106 cells. ChIP was performed with the optimal amount of chromatin as suggested by the protocol for each manufacturer using an antibody for the low abundance NCOA2 protein and a negative control IgG. Following enrichment, qPCR was performed using the ChIP-IT qPCR Analysis Kit in order to normalise the data and allow for direct comparison of the results. Data is expressed as binding events detected per 1000 cells which represents the average of the raw data triplicates adjusted for the amount of chromatin in the reaction, the resuspension volume and the primer efficiency.

Enriched ChIP DNA obtained from the ChIP-IT HS kit is of high quality and ChIP-Seq validated

ChIP-Seq data for the low abundance transcription factor NCOA2. ChIP was performed using the ChIP-IT HS kit, an antibody against the low abundance transcriptional co-activator NCOA2 and chromatin from 3 million MCF-7 cells. ChIP DNA was processed and 30 million sequence tags were generated using the Illumina® Next-Gen sequencing platform. The sequence data was analysed to identify all NCOA2 binding sites in the human genome. The left panel shows multiple binding sites across a 1 million base pair region on chromosome 11. The image in the right panel is zoomed in to show NCOA2 peaks in the promoters of two genes.

Material available for download
ChIP-IT High Sensitivity® kit protocol
High Sensitivity Chromatin Preparation kit protocol
Sonication recommendations for ChIP

ChIP-IT High Sensitivity citations

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