Shop All Denaturing Agents

Urea (Thermo Scientific™)

Thermo Scientific Pierce Urea is ≥99% pure, crystalline urea, a protein denaturant with low UV absorptivity. In addition to increasing solubility of hydrophobic molecules, urea is a general protein denaturant, unfolding proteins and altering their three-dimensional structures.

Features of Thermo Scientific Pierce Urea:

• Any concentration up to 8M urea can be quickly and easily prepared.
• Free from UV absorbing materials
• Free from heavy metal contaminants
• Free from degradation products such as ammonia

Applications:
• Solubilizing proteins from inclusion bodies
• Increasing solubility of hydrophobic peptides and proteins
• Denaturing proteins

Chaotropes, substances that disrupt the structure of water interactions, help to solubilize hydrophobic proteins and peptides for a variety of biological purposes. For example, hydrophobic peptides and proteins may be dissolved in coupling buffer containing either 8 molar guanidine·HCl or 4 molar urea prior to immobilization on AminoLink Plus Support to create an affinity column. However, because the degradation products of guanidine and urea include ammonia, which contains a primary amine that will compete for reaction to the support, urea and guanidine solutions must be prepared with high quality reagents immediately before use.
Consequently, some proteins will be irreversibly altered upon interaction with urea solutions and may lose their binding function. Before any large-scale use of urea, it is best to test a small sample and determine whether the denaturing effects will adversely affect the intended use of the protein.

UltraPure™ SDS Solution, 10% (Invitrogen™)

UltraPure™ 10% SDS Solution is a premixed aqueous solution containing 10% sodium dodecyl sulfate (SDS). SDS is a protein denaturant used in polyacrylamide gel electrophoresis for molecular weight determinations. It also is used to dissociate nucleic acid-protein complexes in DNA extraction protocols, to disrupt cell membranes, and to prepare prehybridization and hybridization solutions. UltraPure™ 10% SDS Solution is filtered and ready to use.

UltraPure™ Urea (Invitrogen™)

UltraPure™ Urea is an effective protein denaturant. In a highly concentrated (6 to 8 M) urea solution, many proteins unfold and separate into their constituent polypeptide chains. UltraPure™ Urea is also used to denature nucleic acids in sequencing gels.

SDS, 20% Solution, RNase-free (Invitrogen™)

Ambion® molecular biology grade, SDS, 20% Solution is supplied in one bottle containing 250 mL. The buffer is certified RNase-free, economical, and ready-to-use. SDS is used as a component of both hybridization and wash buffers for northern and Southern blots and has many other uses around the lab.

8M Guanidine-HCl Solution (Thermo Scientific™)

Thermo Scientific Pierce 8M Guanidine-HCl Solution is an accurately formulated, ready-to-use solution of guanidine hydrochloride, that can be easily diluted and pH-adjusted to any concentration below 8M.

Features of 8M Guanidine-HCl Solution:

• Any concentration below 8M guanidine can be quickly and easily prepared.
• No wasting valuable research time weighing and dissolving guanidine crystals
• Free from UV absorbing materials in the range of 225-300nm
• Free from heavy metal contaminants
• Free from degradation products such as ammonia
• Excellent stability

Applications of Guanidine Hydrochloride:
• Solubilizing proteins from inclusion bodies
• Increasing solubility of hydrophobic peptides and proteins
• Denaturing proteins

Chaotropes, substances that disrupt the structure of water interactions, help to solubilize hydrophobic proteins and peptides for a variety of biological purposes. For example, hydrophobic peptides and proteins may be dissolved in coupling buffer containing either 6M guanidine-HCl or 4M urea prior to immobilization on AminoLink Plus Support to create an affinity column. However, because the degradation products of guanidine and urea include ammonia, which contains a primary amine that will compete for reaction to the support, urea and guanidine solutions must be prepared with high quality reagents immediately before use.

Guanidine is a general protein denaturant, unfolding proteins and altering their three-dimensional structure. Consequently, some proteins will be irreversibly altered upon interaction with guanidine solutions and may lose their binding function. Before any large-scale use of guanidine, it is best to test a small sample and determine whether the denaturing effects will adversely affect the intended use of the protein.

Related Products
Guanidine-HCl

ZOOM™ Thiourea (Invitrogen™)

The ZOOM® IEF Fractionator offers a fast, reliable method to reduce sample complexity, enrich low abundance proteins, and increase the dynamic range of detection. Solution phase isoelectric focusing with the ZOOM® IEF Fractionator provides reproducible separations in three hours. Fractionated samples are ready for further analysis by two dimensional gel electrophoresis (2DE), one dimensional gel electrophoresis (1DE), or two dimensional liquid chromatography/mass spectrometry (2D LC/MS).

The ZOOM® IEF Fractionator System (Figure 1) consists of the following components:

• ZOOM® IEF Fractionator is built for safety, reliability, and durability. It includes easy-to-assemble sample chambers, in-built buffer chambers, buffer trough, removable electrode assembly, and safety lid.
• ZOOM® Disks are pre-cast polyacrylamide gels that eliminate the need for manual preparation and minimize any chance of cross contamination. These immobilized buffered disks are pre-labeled, disposable, and designed for single use, ensuring consistent and reproducible fractionation. They resolve samples into as many as 7 fractions (using 8 disks of specific pH) to six fractions (using six disks of specific pH) from pH 3-12. Simply place each disk between the sample chambers in the ZOOM® IEF Fractionator to allow separation in a specific pH range.
• ZOOM® Reagents are proteomic grade reagents, including ZOOM® 2D Protein Solubilizers urea, thiourea, CHAPS, and ampholytes, that ensure optimal conditions for sample preparation.
• Basic Protein Kit which includes ZOOM® Discs and ZOOM Strips for IEF of basic proteins, focusing buffers, and solubilizer kits.

Sample Chamber and Fractionator Unit Set-up Procedure
Figures 4-6 illustrate assembly of the sample chambers and fractionator unit. The sample chamber and components fit together easily and offer a leak-proof seal. Simply place disks between the sample chambers in the ZOOM® IEF Fractionator to allow separation in a specific pH range. For example, if you want to fractionate proteins between pH 4.6 and 5.4, flank a sample chamber with a ZOOM (r) Disk of pH 4.6 and one of pH 5.4.

Downstream Processing by 2D Gel Electrophoresis
Following fractionation, the separated samples can be subjected to further analysis using narrow or broad range ZOOM® Strips on the ZOOM® IPGRunner™ System. Follow with second dimension analysis using neutral pH NuPAGE® ZOOM® Gels.

Formamide (Deionized) (Invitrogen™)

Ambion formamide has been deionized, is nuclease-free and comes packaged as a liquid in an amber glass bottle under argon. Supplied in one bottle containing 500 g each.Formamide lowers the melting temperature of nucleic acid duplexes and is widely used in applications such as Southern and northern blotting, nuclease protection assays, S1 mapping, and in situ hybridization. For critical applications, it is recommended that the formamide from previously opened bottles be deionized before use.

Appearance: Clear, colorless
Molecular Weight: 45.04
Purity: >99.5%
Quality Control: The product has been tested for RNase and DNase contamination.

Inclusion Body Solubilization Reagent (Thermo Scientific™)

The Thermo Scientific Inclusion Body Solubilization Reagent effectively retrieves recombinant proteins that form insoluble aggregates during bacterial overexpression.

Features of Inclusion Body Solubilization Reagent :

• Achieves the essential first step for subsequent insoluble protein refolding procedures
• One easy- and ready-to-use solution; no preparation needed
• Compatible with Thermo Scientific Pierce Coomassie Plus Protein Assay Kit (Part No. 23236) to determine degree of expression
• Proprietary denaturant can be easily removed by dialysis so that SDS-PAGE can be performed
• Can accommodate additional components such as reducing agents or chelating agents

The Inclusion Body Solubilization Reagent is a ready-to-use aqueous solution of denaturant that helps to dissolve aggregated proteins (inclusion bodies) that have been separated from the soluble fraction of bacterial cell lysates. The reagent is designed to retrieve expressed protein in soluble form after lysis and extraction with reagents such as Thermo Scientific B-PER Bacterial Protein Extraction Reagent. Bacteria are widely used for recombinant protein expression. However, 70%-80% of proteins produced by recombinant techniques in E. coli form inclusion bodies (i.e., protein aggregates). Once these aggregates are formed, it is very difficult to solubilize them. A proprietary denaturant contained in this reagent provides the most effective means for solubilizing aggregated proteins. Additional components, such as a reducing agent and a chelating agent, may be added to the reagent, depending on the particular application.

SDS, 10% Solution, RNase-free (Invitrogen™)

Molecular biology grade, SDS, 10% Solution is supplied in one bottle containing 500 mL. The buffer is certified RNase-free, economical, and ready-to-use. SDS is used as a component of both hybridization and wash buffers for northern and Southern blots and has many other uses around the lab.

Guanidine Thiocyanate (Invitrogen™)

Guanidine Thiocyanate is an ultrapure, molecular biology grade reagent. It is free of detectable nuclease and protease activity and is a strong protein denaturant, as both the guanidinium cation and the thiocyanate anion are chaotropic agents. It is provided in one bottle containing 500 g. It is recommended for isolation of RNA, especially for tissues such as pancreas with high levels of RNase activity. In solution, it is known as guanidinium thiocyanate. Guanidine Thiocyanate is thoroughly tested for contaminating nonspecific endonuclease, exonuclease, and RNase activity.

Specifications:
Appearance: White crystals Molecular Weight: 118.16 Purity:
>99% Stability: Unlimited under dry ambient conditions. Keep in dark container.

UltraPure™ Formamide (Invitrogen™)

UltraPure™ Formamide is commonly used for denaturing nucleic acids for sequencing gel electrophoresis, electron microscopy, and hybridization. A liquid at room temperature, UltraPure™ Formamide is vacuum distilled and packaged under dry nitrogen and can be used as supplied for many applications within three months of date of purchase. Because the breakdown products of formamide degrade nucleic acids, for sensitive applications formamide should be deionized by treatment with a mixed-bed ion-exchange resin immediately before use.

Guanidine Isothiocyanate (Invitrogen™)

UltraPure™ Guanidine Isothiocyanate is a strong protein denaturant when used at high concentrations. It is useful in the isolation of intact RNA from sources rich in RNase such as pancreas (1).

Guanidine-HCl (Thermo Scientific™)

Thermo Scientific Pierce Guanidine-HCl can be easily dissolved and pH-adjusted to any concentration.

Features of Guanidine-HCl:

• No wasting valuable research time weighing and dissolving guanidine crystals
• Free from UV absorbing materials in the range of 225-300nm
• Free from heavy metal contaminants
• Free from degradation products such as ammonia
• Excellent stability

Applications of Guanidine Hydrochloride:
• Solubilizing proteins from inclusion bodies
• Increasing solubility of hydrophobic peptides and proteins
• Denaturing proteins

Chaotropes, substances that disrupt the structure of water interactions, help to solubilize hydrophobic proteins and peptides for a variety of biological purposes. For example, hydrophobic peptides and proteins may be dissolved in coupling buffer containing either 6M guanidine-HCl or 4M urea prior to immobilization on AminoLink Plus Support to create an affinity column. However, because the degradation products of guanidine and urea include ammonia, which contains a primary amine that will compete for reaction to the support, urea and guanidine solutions must be prepared with high quality reagents immediately before use.

Guanidine is a general protein denaturant, unfolding proteins and altering their three-dimensional structure. Consequently, some proteins will be irreversibly altered upon interaction with guanidine solutions and may lose their binding function. Before any large-scale use of guanidine, it is best to test a small sample and determine whether the denaturing effects will adversely affect the intended use of the protein.

Related Products
8M Guanidine-HCl Solution

Hi-Di™ Formamide (Applied Biosystems™)

This is highly deionized formamide used to resuspend samples before electrokinetic injection on capillary electrophoresis systems.

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

ZOOM™ Urea (Invitrogen™)

The ZOOM® IEF Fractionator offers a fast, reliable method to reduce sample complexity, enrich low abundance proteins, and increase the dynamic range of detection. Solution phase isoelectric focusing with the ZOOM® IEF Fractionator provides reproducible separations in three hours. Fractionated samples are ready for further analysis by two dimensional gel electrophoresis (2DE), one dimensional gel electrophoresis (1DE), or two dimensional liquid chromatography/mass spectrometry (2D LC/MS).

The ZOOM® IEF Fractionator System (Figure 1) consists of the following components:

• ZOOM® IEF Fractionator is built for safety, reliability, and durability. It includes easy-to-assemble sample chambers, in-built buffer chambers, buffer trough, removable electrode assembly, and safety lid.
• ZOOM® Disks are pre-cast polyacrylamide gels that eliminate the need for manual preparation and minimize any chance of cross contamination. These immobilized buffered disks are pre-labeled, disposable, and designed for single use, ensuring consistent and reproducible fractionation. They resolve samples into as many as 7 fractions (using 8 disks of specific pH) to six fractions (using six disks of specific pH) from pH 3-12. Simply place each disk between the sample chambers in the ZOOM® IEF Fractionator to allow separation in a specific pH range.
• ZOOM® Reagents are proteomic grade reagents, including ZOOM® 2D Protein Solubilizers urea, thiourea, CHAPS, and ampholytes, that ensure optimal conditions for sample preparation.
• Basic Protein Kit which includes ZOOM® Discs and ZOOM Strips for IEF of basic proteins, focusing buffers, and solubilizer kits.

Sample Chamber and Fractionator Unit Set-up Procedure
Figures 4-6 illustrate assembly of the sample chambers and fractionator unit. The sample chamber and components fit together easily and offer a leak-proof seal. Simply place disks between the sample chambers in the ZOOM® IEF Fractionator to allow separation in a specific pH range. For example, if you want to fractionate proteins between pH 4.6 and 5.4, flank a sample chamber with a ZOOM (r) Disk of pH 4.6 and one of pH 5.4.

Downstream Processing by 2D Gel Electrophoresis
Following fractionation, the separated samples can be subjected to further analysis using narrow or broad range ZOOM® Strips on the ZOOM® IPGRunner™ System. Follow with second dimension analysis using neutral pH NuPAGE® ZOOM® Gels.