CUTANA™

CUTANA™ Uncharged pAG-Tn5 for CUT&Tag

$645.00
SKU: 15-1025
Pack Size: 20 µL
  • Type:  Transposase
  • Mol Wgt:  157 kDa (dimer)
  • Host:  E. coli
  • Epitope Tag:  None

Description

Products in EpiCypher’s IDEA Toolbox (Innovation and Discovery of Epigenetic Applications) offer access to reagents without known or fully defined uses, enabling researchers to explore cutting-edge applications. Due to their novelty and unexplored potential, EpiCypher will engage in limited technical support.

CUTANA Uncharged pAG-Tn5 is a fusion of Proteins A and G to E. coli transposase (Tn5) and is the key enzyme for CUT&Tag [1]. This product is highly purified to remove contaminating E. coli DNA. For superior normalization, as well as antibody validation and reaction monitoring, SNAP-CUTANA™ nucleosome spike-ins (e.g. EpiCypher 19-1002) are recommended. This Tn5 is uncharged and must be loaded with user-designed mosaic adapter DNA prior to use in CUT&Tag. For a charged enzyme that is ready to use, see EpiCypher 15-1017.

Validation Data

Figure 1: Protein gel data
CUTANA pAG-Tn5, Uncharged (1 µg) was resolved via SDS-PAGE and stained with Coomassie blue. The migration and molecular weight of the protein standards are indicated.

Recommended Accessory Products

Item Cat. No.
CUTANA™ Concanavalin A Conjugated Paramagnetic Beads 21-1401
CUTANA™ Rabbit IgG CUT&RUN Negative Control Antibody 13-0042
Histone H3K4me3 Antibody, SNAP-Certified™ for CUT&RUN and ChIP 13-0041
Anti-Rabbit Secondary Antibody for CUTANA™ CUT&Tag 13-0047
Anti-Mouse Secondary Antibody for CUTANA™ CUT&Tag 13-0048
SNAP-CUTANA™ K-MetStat Panel 19-1002
CUTANA™ Non-Hot Start 2X PCR Master Mix for CUT&Tag 15-1018
Magnetic Separation Rack, 0.2 mL Tubes 10-0008
Magnetic Separation Rack, 1.5 mL Tubes 10-0012
CUTANA™ CUT&RUN 8-strip 0.2 mL Tubes 10-0009

Technical Information

Storage
Stable for one year at -20°C from date of receipt. The protein is not subject to freeze/thaw under these conditions.
Formulation
50 mM HEPES-KOH pH 7.2, 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.1% Triton X-100, 50% glycerol

Application Notes

pAG-Tn5 transposomes can be assembled as previously described [1]. In brief, 3.2 µL of a 100 µM equimolar mixture of pre-annealed user-defined Adapter-A and user-defined Adapter-B oligonucleotides should be mixed with 20 µL of 5.5 µM pAG-Tn5 fusion protein dimer. The mixture is then incubated on a rotating platform for 1 hour at room temperature and stored at -20°C. Specific activity definition of the charged pAG-Tn5 is highly recommended before use in CUT&Tag. Due to the confounding variable of user-supplied mosaic adapters, EpiCypher will not engage in protocol troubleshooting for this reagent. For a pre-charged enzyme that is eligible for technical support, see EpiCypher 15-1017.

References

Background References:
[1] Kaya-Okur et al. Nat. Commun. (2019). PMID: 31036827

Documents & Resources

Intellectual Property

US Pat. No. 10,689,643, 11,306,307, EU Pat. No. 3,688,157, 2,999,784 and related patents and pending applications

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