TALON CellThru
His-tagged protein purification from crude cell lysates—TALON CellThru
TALON CellThru is a novel IMAC resin for purifying his-tagged proteins from crude cell lysates, sonicates, and fermentation harvests. This resin uses the same proprietary ligand as our TALON Metal Affinity Resin but has larger beads (300–500 µm) which permit cellular debris to flow through without centrifugation. TALON CellThru Resin captures his-tagged protein directly from crude lysates in one quick step, minimizing protein degradation and generating higher yields of purified protein than conventional strategies.
TALON CellThru Resin is ideal for purifying membrane-bound proteins or multi-protein complexes, which typically pellet with the cell debris during the lysate clarification step and are not available for binding to the resin. Because crude, nonclarified lysates can be applied directly to the resin, you can skip the centrifugation step, allowing membrane-bound proteins and multi-protein complexes to remain in the lysate where they can bind to the resin.
Overview
- Purification from crude cell lysate
- Ideal for purification of membrane-bound protein or multiprotein complexes
- No copurification of proteins
- Low metal ion leakage
- Fast & easy protocol
Applications
Purification of protein from:
- Crude cell lysates
- Sonicates
- Fermentation harvests
SDS-PAGE of TALON CellThru Resin purified proteins
SDS-PAGE of TALON CellThru Resin purified proteins. E. coli BL21 cells were sonicated in TALON wash buffer and run through a TALON CellThru column eluted in 150 mM imidazole. Note that some target protein is trapped in membrane fractions and does not get absorbed on the column. M: molecular weight marker.
Native vs. denaturing purification procedures using TALON resin
Native vs. denaturing purification procedures using TALON resin.
Native purification with TALON resin preserves biological activity of proteins
Native purification with TALON resin preserves biological activity of proteins. Fresh cells (0.5 g) expressing 6xHis-GFPuv were extracted in 5 ml of 50 mM sodium phosphate; 0.3 M NaCl, pH 7.0 Panel A. Elution profile of GFP which was loaded, washed with the same buffer, and eluted with a step gradient of imidazole (150 mM). Panel B. Fractions were analyzed by SDS-PAGE. The fluorescent signal of green fluorescent protein (GFPuv) was completely enriched by TALON Superflow Resin.
Purification of 6xHis-GFPuv under denaturing conditions using TALON resin
Purification of 6xHis-GFPuv under denaturing conditions using TALON resin. The fusion protein was purified in 8 M urea using TALON resin. M=molecular weight markers.
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SDS-PAGE of TALON CellThru Resin purified proteins
SDS-PAGE of TALON CellThru Resin purified proteins. E. coli BL21 cells were sonicated in TALON wash buffer and run through a TALON CellThru column eluted in 150 mM imidazole. Note that some target protein is trapped in membrane fractions and does not get absorbed on the column. M: molecular weight marker.
Native vs. denaturing purification procedures using TALON resin
Native vs. denaturing purification procedures using TALON resin.
Native purification with TALON resin preserves biological activity of proteins
Native purification with TALON resin preserves biological activity of proteins. Fresh cells (0.5 g) expressing 6xHis-GFPuv were extracted in 5 ml of 50 mM sodium phosphate; 0.3 M NaCl, pH 7.0 Panel A. Elution profile of GFP which was loaded, washed with the same buffer, and eluted with a step gradient of imidazole (150 mM). Panel B. Fractions were analyzed by SDS-PAGE. The fluorescent signal of green fluorescent protein (GFPuv) was completely enriched by TALON Superflow Resin.
Purification of 6xHis-GFPuv under denaturing conditions using TALON resin
Purification of 6xHis-GFPuv under denaturing conditions using TALON resin. The fusion protein was purified in 8 M urea using TALON resin. M=molecular weight markers.
