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- Cat. No.:77393-514
- Contents:2×50 ml binding buffer; 2×50 ml wash buffer; 1×50 ml elution buffer; 1×10 ml 100 Ni-INDIGO resin; 5×MIDI spin columns; 50×collection tubes; 1×500 mM DTT (add 1.016 ml ddH₂O); 1×500 mM EDTA (add 0.866 ml ddH₂O)
The HisCube His-tag protein purification MIDI kit is our way to supply your laboratory with a convenient method for high-quality, small-scale His-tag protein purifications. We put all of our experience in His-tag purifications into this Kit to provide a protocol that is easy to follow.
- Usage: Enrichment of phosphopeptides
- Specificity: Affinity to phosphorylated biomolecules
- Bead ligand: Ti-NTA
- Bead size: 30 µm
- Filling quantity: Delivered as a 25% suspension
- Chelator stability: Stable in buffer containing 10 mM DTT and 1 mM EDTA
- pH stability: 2 to 14
- Other stabilities: 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile
The HisCube Ni-INDIGO MIDI Kit contains our in-house developed PureCube 100 Ni-INDIGO agarose resin and our PureCube 1-Step Batch MIDI batch spin tubes.
INDIGO's most significant advantage is its tolerance towards reducing and chelating agents. These can be used to great help in His-tag purifications, but traditional His-tag affinity beads do not tolerate them. EDTA is a chelator often used in eukaryotic cell buffers because it has protease-inhibiting functions. DTT can dissolve protein aggregates that could hinder the access of the purification beads to the His-tagged proteins. However, thanks to INDIGO, both substances can be used now. Therefore, they are part of the kit.