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Thermo Scientific™ Pierce™ Protein Refolding Kit

Catalog No. PI89867
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100 Reactions
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PI89867 100 Reactions
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Catalog No. PI89867 Supplier Thermo Scientific™ Supplier No. 89867

Description

Refold recombinant proteins that have been denatured and solubilized from inclusion bodies by determining optimal buffer conditions using this reagent matrix strategy.

Thermo Scientific™ Pierce Protein Refolding Kit includes high-purity reagents and detailed instructions for using a matrix strategy to determine optimal buffer conditions for refolding recombinant proteins that have been denatured and solubilized from inclusion bodies.

The kit contains the essential reagents and complete strategy for determining optimal buffer conditions to refold denatured recombinant proteins to restore native structure and function. Nine base refolding buffers form a matrix that includes a range of strong and weak denaturant conditions for the suppression of protein aggregation. The supplied additives are used as additional matrix factors, depending on the protein type being refolded. Buffer components are examined at three concentration levels, allowing a wide spectrum of folding conditions to be tested within one experiment. The adjustable design allows matrix conditions to be tailored to the target protein, preventing sample waste and unnecessary analysis, while maximizing refolding yields. The Pierce Protein Refolding Kit is accompanied by a comprehensive Refolding Guide with details on isolating, solubilizing and purifying inclusion bodies; optimizing refolding conditions; and analyzing refolding yields.

Highlights:

  • Robust – conditions and components examined are limited to those having the most significant and general utility as folding buffers

  • Convenient – three-level matrix design significantly reduces the amount of secondary optimization required and increases the ease of data interpretation

  • Adjustable matrix format – allows refolding experiments to be customized to the target protein; known positive and negative interactions between buffer components are addressed, minimizing unnecessary analyses

  • High-purity reagents – reagents are formulated using stringent standards so that consistent results are attained

Includes:

Nine different Base Refolding Buffers (10mL each), convenient amounts of seven additives, and a comprehensive Refolding Guidebook

Compatible with:

Thermo Scientific Inclusion Body Solubilization Reagent (Part No. 78115)

Specifications

Product Type Protein Refolding Kit
Content And Storage Upon receipt store at -20°C. Product may be stored at 4°C for up to two months. Product is shipped on dry ice.
Form Liquid, Powder
Product Line Pierce
Reagent Type Chaotrope, Denaturant
Quantity 100 Reactions

Frequently Asked Questions (FAQs)

Can you provide suggestions on how to refold protein following purification?

Please read our suggestions below:

-Maintain low protein concentrations 10-50 µg/mL.
-Disulfide bonds help to stabilize native proteins; add a redox pair such as GSH (reduced glutathione)
-GSSG (oxidized glutathione) at a ratio of 10:1 with GSH concentration of 2-5 mM. Such a redox pair is helpful to create an oxidizing potential to break and build new S-S bonds during the folding process.
-Remove denaturants slowly by dilution or dialysis; glycine (50 mM, pH 9.0, 5 mM EDTA) helps to solubilize the proteins. If GuHCl (guanidinium HCl) is used, add 2 M urea because urea helps to stabilize the protein folding, too.
-Add detergents at a very low concentration such as 0.1-0.5% NP-40 or 0.005% (v/v) Tween 20.
-Include co-solvents to stabilize the proteins such as glycerol (5-20%) or PEG 8000 or glucose or sucrose. (10 %)
-Certain anions (e.g., phosphate or sulfate) or cations (e.g., MES or HEPES) have positive effects, too; include salt and maintain a neutral pH such as 100 mM KCl, or 150-500 mM NaCl, 2 mM MgCl2.
-Avoid protein degradation by adding protease inhibitors such as 0.5 mM PMSF, 0.005-2 µg/mL aprotinin, 2 µg/mL Pepstatin, or 2-5 µg/mL leupeptin.
-Dialysis of a phosphate buffer when against calcium will result in a calcium phosphate precipitate. If CaCl2 is needed for subsequent enterokinase digestion (10 mM Tris pH 8, 10 mM CaCl2), remember to add CaCl2 after dialysis is complete.
Alternatively, you can use the Pierce Protein Refolding kit (cat. No. 89867)

For Research Use Only. Not for use in diagnostic procedures.