LYTAG Two-Phase Purification system is a protein purification system based on the use of two aqueous components. The method relies on the affinity of the protein tag LYTAG for one of the two-phase components, allowing recombinant protein separation and purification from cellular extracts or culture media. In the procedure, the LYTAG-fused protein is retained in one of the aqueous phases while most of the undesired proteins can be removed by simply discarding the opposite phase. After replenishing the system with fresh phase, the protein of interest can be easily recovered in it, with high purity, by reversing its localization with the addition of choline, the specific LYTAG ligand.
This system is particularly well suited for industries and laboratory specialized in protein separation and purification, as it is simple, cost efficient, time saving and highly versatile for scaling up protein purification process, representing a convenient alternative to solid resins.
- Quick method. Separation can be completed within few minutes, minimizing the effects of proteases. It is especially convenient when large volumens (>10 ml) of cellular extracts or culture media need to be processed (usually requiring prolonged flow times when using chromatography columns).
- Easy and inexpensive protein purification process.
- No special equipment requirements.
- It offers, like every aqueous two-phase systems, mild conditions in the separation of labile proteins. Purification can be easy and safely performed at low temperatures, requiring only a refrigerated centrifuge and an ice water bath.
- Scalable method.
- High purification efficiency (>95% purity), comparable to the use of solid matrixes.
- Optimum performance in downstream fermentation processes.
- Good alternative to conventional solid resins.
LYTAG 2-PHASE Purification System
- 1M Salicylate (Inducer), 25ml
- pALEX2a, 8μg
- pALEX2b, 8μg
- pALEX2c, 8μg
- pALEX-Ca-GFP (expression and purification control plasmid, 8μg)
- E. Coli REG 12, stab
- Top solution
- Lower solution
- LYTAG Two-Phase Wash Buffer
- Elution Buffer
AB-3237 Mouse AntiHis-tag (monoclonal) N-terminal HisTag 100 μl RS-3322 Empty Purification Columns (2.5 ml) 1 units RS-3396 Empty Purification Columns (2.5 ml) 10 units RS-3414 Empty Purification Columns (5 ml) 1 units RS-3415 Empty Purification Columns (10 ml) 1 units RS-4700 Lower Solution (LYTAG 2-Phase Purification System) 20 ml RS-4701 Top Solution (LYTAG 2-Phase Purification System) 40 ml RS-4702 LYT 2-Phase Wash (2x) (LYTAG 2-Phase Purification System) 125 ml RS-4740 Top Solution (LYTAG 2-Phase Purification System) 80ml RS-4755 Choline Chloride 3M (LYTAG 2-Phase Purification System) 10 ml RS-4756 Choline Chloride 3M (LYTAG 2-Phase Purification System) 25 ml