Profinity eXact™ Purification and Tag Cleavage Consumables

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Overview

Profinity eXact™ Fusion-Tagged Protein Purification
SDS-PAGE analysis of a purification of Profinity eXact fusions to a 26 kD Green Fluorescent Protein (GFP) and to a 40 kD maltose binding protein (MBP).

SDS-PAGE analysis of a purification of Profinity eXact fusions to a 26 kD green fluorescent protein (GFP) and to a 40 kD maltose binding protein (MBP). Crude E. coli lysate containing the expressed proteins (2 ml) was loaded onto a Bio-Scale Mini Profinity eXact 1 ml cartridge on a BioLogic DuoFlow™ system with bind/wash buffer at 1 ml/min. The cartridge was washed with 10 column volumes (CV) of the same buffer at 1 ml/min. Proteins were eluted with 3 CV sodium fluoride buffer at room temperature and at 0.1 ml/min for 30 min. Total purification time required to generate the tag-free proteins without addition of protease was approximately 60 min.

Stability and Reusability of Profinity eXact™ Purification Resin
Overlaid chromatograms of MBP purification

Stability and reusability of Profinity eXact purification resin. Overlaid chromatograms of MBP purification with a 1 ml Bio-Scale Mini Profinity eXact cartridge on the BioLogic DuoFlow system. Binding capacities for cycles 1 to 5 ranged from 3.7 mg/ml to 3.3 mg/ml.



Profinity eXact consumables include Profinity eXact purification resin, Profinity eXact mini spin columns, Bio-Scale Mini Profinity eXact cartridges, and the Profinity eXact expression and purification starter kit.

Profinity eXact Purification Resin

The Profinity eXact purification resin consists of a highly engineered subtilisin protease conjugated to an agarose-based matrix. Purity of the eluted protein using this purification resin is typically higher than that for other affinity-tag systems. This is due to the specific recognition of subtilisin for its prodomain sequence. The resin can be packed into different column formats, including low- to medium-pressure columns, gravity-flow columns, and mini spin columns, offering users added purification flexibility.

Additional advantages of Profinity eXact purification resin include:

  • Purification and processing of fusion-tagged proteins in a single-step
  • On-column cleavage in as little as 30 min
  • No protease addition required
  • Precise cleavage at N-terminus to generate native protein sequence

The resin is supplied in 100 mM sodium phosphate, pH 7.2, containing 0.02% sodium azide. It is also available prepacked in 1 ml and 5 ml cartridges, and in mini spin columns.

Profinity eXact Mini Spin Columns

Profinity eXact mini spin columns are ideal for rapid expression screening and optimization of affinity-tag purification protocols. These single-use columns contain 0.1 ml settled Profinity eXact resin and all necessary plasticware for binding, washing, and elution steps. Profinity eXact mini spin columns have been designed to fit in all major benchtop centrifuges. The mini spin columns are available separately or in kits:

  • Profinity eXact mini spin column ten-packs (each column contains 0.1 ml settled resin) with collection tubes for binding, washing, and elution steps
  • Profinity eXact mini spin purification starter kit (includes ten mini spin columns, collection tubes, control protein lysate, bacterial lysis reagent, bind/wash buffer, and elution buffer)
  • Profinity eXact expression and purification starter kit (includes Profinity eXact mini spin purification starter kit and Profinity eXact cloning and expression starter kit)

Bio-Scale Mini Profinity eXact Cartridges

These 1 ml and 5 ml cartridges can be used with BioLogic chromatography systems, the Profinia protein purification system, a peristaltic pump, or with appropriate column adaptors with any other chromatography system. These cartridges are prepacked with Profinity eXact purification resin.

Profinity eXact Expression and Purification Starter Kit

The Profinity eXact expression and purification starter kit can be used to easily evaluate Profinity eXact fusion-tag technology and is suitable for new purifications requiring tag removal and for purifications where cleavage has resulted in inferior results. The Profinity eXact expression and purification starter kit includes two kits: the Profinity eXact cloning and expression starter kit for the cloning and expression of a target gene using pPAL vectors and competent cells, and the Profinity eXact mini spin purification starter kit for single-step purification and on-column cleavage of the tagged protein.

Functional ligand Subtillisin protease, 27.8 kD
Base bead 6% agarose bead
Form 50% suspension in 100 mM sodium phosphate, pH 7.2, 0.02% sodium azide
Practical size range 60–160 µm
Dynamic binding capacity >3 mg tag-free protein/ml resin
Recomended linear flow rate 1,000 cm/hr at 25°C
pH stability 2–13
Chemical compatibility Common reagents including detergents, reducing agents, buffering agents, and additives.
Storage 4°C
Shelf life in 20% ethanol >1year at 4°C
Operational Temperature 4–40°C
Profinity eXact Purification Resin

156-3005
10 ml, purification resin supplied in 100 mM sodium phosphate, pH 7.2 with 0.02% sodium azide

List Price:   $712.00
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Profinity eXact Mini Spin Columns

156-3007
Pkg of 10 mini spin columns packed with 0.1 ml Profinity eXact resin, includes 50 capped, collection tubes

List Price:   $213.00
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Bio-Scale Mini Profinity eXact Cartridges

732-4646
Pkg of 2, 1 ml, prepacked affinity column, 40 mm x 5.6 mm, max. pressure 45 psi, for use with BioLogic chromatography and Profinia systems

List Price:   $274.00
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Bio-Scale Mini Profinity eXact Cartridges

732-4647
Pkg of 4, 1 ml, prepacked affinity column, 40 mm x 5.6 mm, max. pressure 45 psi, for use with BioLogic chromatography and Profinia systems

List Price:   $479.00
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Bio-Scale Mini Profinity eXact Cartridge

732-4648
Pkg of 1, 5 ml, prepacked affinity column, 40 mm x 12.6 mm, max. pressure 45 psi, for use with BioLogic chromatography and Profinia systems

List Price:   $515.00
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Profinity eXact Expression and Purification Starter Kit

156-3008
Kit includes 1 Profinity eXact cloning and expression starter kit (20 reactions) and 1 Profinity eXact mini spin purification starter kit (10 spin columns)

List Price:   $965.00
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Profinity eXact Mini Spin Purification Starter Kit

156-3006
Mini purification starter kit includes 10 prepacked spin columns, collection tubes, 1 vial lyophilized lysate, lysis reagent and purification buffers

List Price:   $625.00
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Number Description Options
6334 Tips and Tricks for Successful Preparative Chromatography and Protein Purification, Rev B Click to download
5652 Profinity eXact Fusion-Tag System Performs On-Column Cleavage and Yields Pure Native Protein From Lysate in Less Than an Hour, Rev A Click to download [ Add to Cart (Free) ]
5656 Purification of Tag-Free Recombinant Proteins Using the Profinity eXact Fusion-Tag System, Rev B Click to download [ Add to Cart (Free) ]
5668 Profinity eXact Fusion-Tag System Brochure, Rev A Click to download [ Add to Cart (Free) ]
5646 Profinity eXact Cloning and Expression Kits, PIS, Rev B Click to download [ Add to Cart (Free) ]
5655 Profinity eXact Resin Product Information Sheet, Rev A Click to download [ Add to Cart (Free) ]
5811 Profinity eXact Fusion-Tag System FAQs, Rev A Click to download
5898 Using the Profinity eXact Fusion-Tag System: Strategies for Success, Rev A Click to download
10011161 BL21(DE3) Chemi-Competent Expression Cells Manual, Rev A Click to download
10011164 Bio-Scale Mini Profinity eXact Cartridges Manual, Rev B Click to download
10011260 Profinity eXact Protein Purification System Manual, Rev A Click to download [ Add to Cart (Free) ]
5772 Periplasmic Expression and Purification of Recombinant Proteins from E.coli Using the Profinity eXact Fusion-Tag System, Rev A Click to download [ Add to Cart (Free) ]
5928 Application of the Profinity eXact Fusion-Tag System to Eukaryotic Expression Systems Tech Note, Rev B Click to download [ Add to Cart (Free) ]