Key features and details
- Expression system: Escherichia coli
- Active: Yes
- Suitable for: ChIC/CUT&RUN, ChIC/CUT&RUN-seq
Product nameChIC/CUT&RUN pAG-MNase
pAG-MNase enzyme for performing ChIC/CUT&RUN experiments. Biological activity determined by using pAG-MNase in the ChIC/CUT&RUN experiment on 1 X105 HeLa cells with ab188408 Anti-CTCF antibody followed by sequencing. Click here for the full list of ChIC/CUT&RUN-seq validated antibodies.
pAG-MNase is supplied in 10 mM Tris HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, and 50% glycerol. Use 2 µl pAG-MNase per 50 µl ChIC/CUT&RUN reaction (25X dilution).
Number of reactions:
100 µl vial (50 reactions)
500 µl vial (250 reactions)
Expression systemEscherichia coli
Protein lengthFull length protein
Additional sequence informationNuclease
DescriptionRecombinant MNase protein
Our Abpromise guarantee covers the use of ab285373 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ChIC was first described by the Laemmli group in 2004 (1). The ChIC method involves cleavage of double-stranded DNA (1). The antibody directed pAG-MNAse fusion protein (ab285373) results in 100 – 200 bp resolution mapping of DNA-bound proteins (1, 2). ChIC/CUT&RUN-seq allows chromatin immunoprecipitation to be performed on fewer than 100 cells for histone modifications and ~1000 cells for transcription factors compared to ChIP-seq which generally needs >106 cells (1, 3). Another advantage of ChIC/CUT&RUN-seq over ChIP-seq is an ~10-fold lower sequencing depth (1, 3). Abcam provides an increasing number of ChIC/CUT&RUN-seq validated antibodies to meet the needs of this emerging technique.
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Preparation and Storage
Stability and Storage
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Information available upon request.
This product is an active protein and may elicit a biological response in vivo, handle with caution.
Figure 1: Protein gel of pAG-MNAse
pAG-MNase ab285373 (16 µg per lane) was analysed by SDS-PAGE and stained using Coomassie Blue. The molecular weights are outlined alongside the target protein.
Figure 2: Size distribution of cleaved chromatin
ChIC/CUT&RUN was performed with ab285373 pAG-MNase (1:25 dilution) and Anti-CTCF antibody (ab188408). Library preparation was performed and the size of excised DNA fragments was analysed by an Agilent 4200 TapeStation. Enriched cleaved DNA mononucleosomes are observed at ~300 bp.
Figure 3: ChIC/CUT&RUN sequencing - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
ChIC/CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373, 1x105 HeLa cells and 5µg of ab188408 [EPR18253]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The ChIP data was conducted on chromatin prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 1x107 HeLa cells and 4 µg of ab188408 [EPR18253]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Figure 4: ChIC/CUT&RUN sequencing - Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade (ab213224)
ChIC/CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373, 1x105 HeLa cells and 2µg of ab213224 [EPR20551-225]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The ChIP data was conducted on chromatin prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 1x107 HeLa cells and 4 µg of ab213224 [EPR20551-225]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab285373 has not yet been referenced specifically in any publications.