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CUTANA™ pAG-MNase for ChIC/CUT&RUN Assays

ChIC, CUT&RUN 20 X

Size Distribution of Released Chromatin:CUT&RUN was performed using CUTANA™ pAG-MNase (1:20 dilution) with 0. (CUTANA™ pAG-MNase for ChIC/CUT&RUN Assays)

Protein Gel Data: CUTANA™ pAG-MNase (1 μg) was resolved via SDS-PAGE and stained with Coomassie blue. (CUTANA™ pAG-MNase for ChIC/CUT&RUN Assays)

CUT&RUN Data:Sequencing tracks obtained using CUTANA™ pAG-MNase. (CUTANA™ pAG-MNase for ChIC/CUT&RUN Assays)

Summary of the CUT&RUN protocol using a primary and secondary antibody (left). (CUTANA™ pAG-MNase for ChIC/CUT&RUN Assays)

5 images

(4 references)

Catalog No. ABIN6950951

Quick Overview for CUTANA™ pAG-MNase for ChIC/CUT&RUN Assays (ABIN6950951)

Target

Nuclease

Application

Chromatin Immunocleavage (ChIC), Cleavage Under Targets and Release Using Nuclease (CUT&RUN)

Host

Escherichia coli (E. coli)

Purpose

This construct of Protein A and G fused to Micrococcal Nuclease is useful in performing CUT&RUN.

The pAG-MNase fusion enzyme may be used in conjunction with any of our CUT&RUN Product Sets.

Brand

CUTANA™

Characteristics

Recombinantly produced in E. coli, CUTANA™ pAGMNase for ChIC/CUT&RUN Workflows is a fusion of Proteins A and G to Micrococcal Nuclease. This construct is useful in performing Chromatin Immunocleavage (ChIC) and Cleavage Under Targets and Release Using Nuclease (CUT&RUN). CUTANA pAG-MNase contains a C-terminal 6xHis epitope tag.
Application Notes

This product is sufficient to perform 50/250 CUT&RUN reactions.
Recommended use: 2.5 μL of the supplied enzyme into a 50 μL CUT&RUN reaction (20X dilution).

Comment
CUTANA pAG-MNase, the essential reagent for ChIC/CUT&RUN workflows:
- First-in-class commercial product for ChIC/CUT&RUN assays
- Optimized fusion of Proteins A and G with Micrococcal Nuclease (pAG-MNase) enables direct compatibility with a broad range of antibody isotypes
- 50 and 250 reaction pack sizes, enabling greater experimental throughput
Restrictions

For Research Use only
Concentration

20 X

Buffer

Provided as a 20X stock in 10 mM Tris HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, and 50 % glycerol.

Handling Advice

Avoid freeze-thaw cycles

Storage

-20 °C

Storage Comment

Stable for one year at -20°C from date of receipt. The protein is not subject to freeze/thaw under these conditions.

Expiry Date

12 months
Meers, Bryson, Henikoff, Henikoff: "Improved CUT&RUN chromatin profiling tools." in: eLife, Vol. 8, (2019) (PubMed).

Skene, Henikoff, Henikoff: "Targeted in situ genome-wide profiling with high efficiency for low cell numbers." in: Nature protocols, Vol. 13, Issue 5, pp. 1006-1019, (2019) (PubMed).

Skene, Henikoff: "An efficient targeted nuclease strategy for high-resolution mapping of DNA binding sites." in: eLife, Vol. 6, (2018) (PubMed).

Schmid, Durussel, Laemmli: "ChIC and ChEC; genomic mapping of chromatin proteins." in: Molecular cell, Vol. 16, Issue 1, pp. 147-57, (2004) (PubMed).
Target

Nuclease

Molecular Weight

44 kDa

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