CUTANA™ Fluorescent pAG-Tn5 for CUT&Tag
54 in stock
Powered by BiozProducts 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 Fluorescent pAG-Tn5 is a fusion of Proteins A and G to E. coli transposase (Tn5), the key enzyme for CUT&Tag [1]. The Mosaic A and Mosaic B adapters loaded in this enzyme bear a 5’ Cy5 fluorescent dye to enable multiomic visualization and mapping of chromatin features. This product is highly purified to remove contaminating E. coli DNA. For an enzyme charged with non-fluorescent adapters that can be used in standard CUT&Tag, see EpiCypher 15-1017.
Intellectual Property
Background References:
[1] Kaya-Okur et al. Nat. Commun. (2019). PMID: 31036827
Figure 1: DNA charging data
CUTANA Fluorescent pAG-Tn5 for CUT&Tag (0.5 µg) was resolved via Native PAGE alongside uncharged pAG-Tn5 (EpiCypher 15-1025) and charged pAG-Tn5 (EpiCypher 15-1017). The gel was imaged and Cy5 emission was observed at 670 nm (right), stained with ethidium bromide, and imaged again at 605 nm. The observed shift indicates successful loading of the Tn5, and fluorescent imaging confirms the presence of the dye.
Figure 2: Size distribution of released chromatin
Figure 3: CUT&Tag data
Figure 4: Protein gel data
- Type: Transposase
- Mol Wgt: 192 kDa
- Host: E. coli
- Epitope Tag: None
50 mM HEPES-KOH pH 7.2, 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.1% Triton X-100, 50% glycerol
Tn5ME-A: 5’-[Cy5]TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3’
Tn5ME-B: 5’-[Cy5]GATTCGTGGGCTCGGAGATGTGTATAAGAGACAG-3’
Tn5ME-rev: 5’-[phos]CTGTCTCTTATACACATCT-3’
Application Notes
A 20X dilution of this reagent is sufficient for a standard CUT&Tag reaction following the EpiCypher CUTANA™ Direct-to-PCR CUT&Tag protocol, indicating that addition of the fluorescent dye does not prevent Tn5-mediated tagmentation of chromatin (see Figures 2 & 3). However, application-specific optimization will be necessary for use in microscopy. Due to the novelty of this reagent, EpiCypher will not engage in application tech support.
| Item | Cat. No. |
|---|---|
| Anti-Rabbit Secondary Antibody for CUTANA™ CUT&Tag | 13-0047 |
| Anti-Mouse Secondary Antibody for CUTANA™ CUT&Tag | 13-0048 |
| CUTANA™ Non-Hot Start 2X PCR Master Mix for CUT&Tag | 15-1018 |
| Histone H3K4me3 Antibody, SNAP-Certified™ for CUT&RUN and ChIP | 13-0041 |
| CUTANA™ Rabbit IgG CUT&RUN Negative Control Antibody | 13-0042 |
