CUTANA™ Uncharged 6xHis-pAG-Tn5 for CUT&Tag
Catalogue Number: 15-1028-EPC
| Manufacturer: | EpiCypher |
| Shelf Life: | 12 months |
| Type: | Other Enzymes |
| Host Cell: | E. coli|E. coli |
| Shipping Condition: | Blue Ice |
| Unit(s): | 75 ul |
Description
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 6xHis-pAG-Tn5 is a fusion of proteins A and G to E.coli transposase (Tn5), the key enzyme for CUT&Tag [1]. The fusion protein contains a polyhistidine tag (6xHis) to enable applications that require affinity purification steps (for an untagged version of uncharged pAG-Tn5, see EpiCypher 15-1025). Uncharged Tn5 must be loaded with user-designed mosaic adapter DNA prior to use in CUT&Tag (see reference [1] and Application Notes). His-pAG-Tn5 enables purification of differentially barcoded antibody-Tn5 complexes such as those described in MulTI-Tag (Multiple Target Identification by Tagmentation) and multi-CUT&Tag workflows to simultaneously interrogate multiple chromatin proteins in a single CUT&Tag reaction [2,3]. These approaches have been applied to map histone post-translational modifications and RNA Polymerase II down to single cell resolution to define cell-specific epigenetic landscapes.
Additional Text
Protein Tag
6xHIS Tag
Storage Note
Stable for one year at -20°C from date of receipt. The protein is not subject to freeze/thaw under these conditions.
Application Notes
pAG-Tn5 transposomes can be assembled as previously described [1]. In brief, 3.2 µL of an equimolar mixture of pre-annealed user-defined Adapter-A and user-defined Adapter-B oligonucleotides (50 µM each, 100 µM total adapter DNA) should be mixed with 70 µL of 1.5 µM pAG-Tn5 fusion protein dimer (a 3:1 molar ratio of adapter DNA to pAG-Tn5 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.
Preparation
75 µL of enzyme in a screw cap plastic vial
15-1028 Datasheet
SUPPORT
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PRODUCT
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DELIVERY
we offer free delivery to UK universities and non profit organisations