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Detergents for Protein Solubilization |
Detergents are essential for protein extraction by aiding the solubilization of proteins from biological samples. Also known as surfactants, detergents lower the surface tension of a liquid and the tension between two liquids (1). They are used for protein solubilization to efficiently extract and dissolve proteins from biological samples for further analysis and experimentation. They are essential components used in cell lysis buffers, for protein solubilization procedures, in wash buffers for ELISA, and various other protein research methods. The use of appropriate detergents also enables the extraction of proteins that are difficult to isolate (membrane or nuclear proteins) by disrupting and dissolving the structure of the protein, which can result in higher proteins yields (2).
Thermo Fisher Scientific offers a variety of detergents specifically designed for protein solubilization including ionic and non-ionic, as well as denaturing and non-denaturing detergents. These detergents play a crucial role in various laboratory processes such as cell lysis, electrophoresis and blotting buffers, protein purification, and structural studies. Choose from our comprehensive range of detergent based on your specific protein solubilization needs.
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Choose your optimal protein solubilization detergent
Non-ionic detergents are milder and less denaturing than ionic detergents. These detergents are commonly used in biochemistry and molecular biology protocols to solubilize proteins without disrupting their structure or function, making them ideal for maintaining the functionality of proteins. They are also used for protein extraction when maintaining native protein-protein interactions is essential for downstream applications such as enzyme assays or immunoassays (see Figure 1). Additionally, non-ionic detergents can be used in various other laboratory applications such as western blotting or immunoprecipitation. These types of detergents do not carry an electrical charge in solution.
| NP-40 | Tween-20 | Tween-80 | Triton X-100 | Triton X-114 | Brij-35 | Brij-58 | |
| Properties | Good for isolating cytoplasmic proteins but not nuclear proteins | Cell lysis and protein isolation for ELISA, Western Blotting, and other immunoassay PBS or TBS wash buffers | Commonly used for cell lysis buffers for performing various protein analysis methods Triton X-114 can be used to separate hydrophobic and hydrophilic membrane proteins in certain conditions | Commonly used for cell lysis buffers Brij-35 can be used as a surfactant in various HPLC applications | |||
| Dialyzable | No | No | No | No | No | No | No |
| Aggregation number* | 149 | -- | 60 | 140 | -- | 40 | 70 |
| MW | 617 | 1,228 | 1,310 | 647 | 537 | 1,225 | 1,120 |
| Micelle MW | 90,000 | -- | 76,000 | 90,000 | -- | 49,000 | 82,000 |
| CMC % w/v (mM) | 0.0179% (0.29) | 0.0074% (0.06) | 0.0016% (0.012) | 0.0155% (0.24) | 0.0113% (0.21) | 0.1103% (0.09) | 0.0086% (0.08) |
| Cloud point (°C) | 80 | 95 | -- | 64 | 23 | >100 | >100 |
| Available size(s) | 6 x 10 mL 50 mL 500 mL | 6 x 10 mL 50 mL 250 mL 500 mL | 6 x 10 mL 50 mL 500 mL | 6 x 10 mL 50 mL 250 mL | 6 x 10 mL | 6 x 10 mL 50 mL 500 mL | 6 x 10 mL |
* Aggregation number is the number of molecules per micelle
Ionic detergents are strong solubilizing agents that cause total membrane disruption by denaturing proteins by breaking protein-protein interactions and destroying protein activity and function. They are often used in biochemistry and molecular biology for a variety of applications, such as cell lysis and protein electrophoresis, and also for protein solubilization, disrupting protein-protein interactions, and membrane protein isolation. These types of detergents contain charged hydrophilic groups in their structure and are classified into cationic detergents (positively charged), anionic detergents (negatively charged), or zwitterionic detergents (both positive and negative changes) (see Figure 1).
| Sodium deoxycholate | SDS (sodium dodecyl sulfate), C12 | SDS (sodium dodecyl sulfate), Lauryl | |
| Properties | Anionic Useful for disrupting and dissociating protein interactions Frequently used in cell lysis buffers (RIPA buffer) | Anionic Strong lysis agent that works on a majority of cell types Not suitable for sensitive protein extraction due to its denaturing properties | |
| Popular for protein electrophoresis and solubilization Eliminate most all secondary chain lengths except C12 | Popular for denaturing polyacrylamide gel electrophoresis (SDS-PAGE) | ||
| Dialyzable | Yes | No | |
| Aggregation number* | 5 | 62 | |
| MW | 414.6 | 288.5 | |
| Micelle MW | 2,000 | 18,000 | |
| CMC % w/v (mM) | 0.083 to 0.249% (2-6) | 0.17-0.23% (6-8) | |
| Cloud point (°C) | Unknown | >100 | |
| Available size(s) | 25 g 5 g | 500 g | 100 g |
* Aggregation number is the number of molecules per micelle
Zwitterionic detergents can solubilize proteins without denaturing them, allowing proteins to maintain their native structure and function. They are used in biological and biochemical research, particularly for the extraction, solubilization, and stabilization of membrane proteins. These types of detergents are unique from ionic and non-ionic detergents, in that they contain both positively and negatively charged functional groups and have no net charge.
| CHAPS | CHAPSO | |
| Properties | Zwitterionic Mild lysis agent effective for the lysis of cultured mammalian cells Suitable for protecting the native state of proteins and maintaining protein activity | |
| Dialyzable | Yes | Yes |
| Aggregation number* | 10 | ~11 |
| MW | 615 | 630.9 |
| Micelle MW | 6,149 | Unknown |
| CMC % w/v (mM) | 0.49-0.62% (8-10) | 0.5% (8) |
| Cloud point (°C) | >100 | Unknown |
| Compatible with Detergent Removal Resin | Yes | No |
| Available size(s) | 5 g 100 g | 1 g 5 g |
* Aggregation number is the number of molecules per micelle
Non-ionic detergents are milder and less denaturing than ionic detergents. These detergents are commonly used in biochemistry and molecular biology protocols to solubilize proteins without disrupting their structure or function, making them ideal for maintaining the functionality of proteins. They are also used for protein extraction when maintaining native protein-protein interactions is essential for downstream applications such as enzyme assays or immunoassays (see Figure 1). Additionally, non-ionic detergents can be used in various other laboratory applications such as western blotting or immunoprecipitation. These types of detergents do not carry an electrical charge in solution.
| NP-40 | Tween-20 | Tween-80 | Triton X-100 | Triton X-114 | Brij-35 | Brij-58 | |
| Properties | Good for isolating cytoplasmic proteins but not nuclear proteins | Cell lysis and protein isolation for ELISA, Western Blotting, and other immunoassay PBS or TBS wash buffers | Commonly used for cell lysis buffers for performing various protein analysis methods Triton X-114 can be used to separate hydrophobic and hydrophilic membrane proteins in certain conditions | Commonly used for cell lysis buffers Brij-35 can be used as a surfactant in various HPLC applications | |||
| Dialyzable | No | No | No | No | No | No | No |
| Aggregation number* | 149 | -- | 60 | 140 | -- | 40 | 70 |
| MW | 617 | 1,228 | 1,310 | 647 | 537 | 1,225 | 1,120 |
| Micelle MW | 90,000 | -- | 76,000 | 90,000 | -- | 49,000 | 82,000 |
| CMC % w/v (mM) | 0.0179% (0.29) | 0.0074% (0.06) | 0.0016% (0.012) | 0.0155% (0.24) | 0.0113% (0.21) | 0.1103% (0.09) | 0.0086% (0.08) |
| Cloud point (°C) | 80 | 95 | -- | 64 | 23 | >100 | >100 |
| Available size(s) | 6 x 10 mL 50 mL 500 mL | 6 x 10 mL 50 mL 250 mL 500 mL | 6 x 10 mL 50 mL 500 mL | 6 x 10 mL 50 mL 250 mL | 6 x 10 mL | 6 x 10 mL 50 mL 500 mL | 6 x 10 mL |
* Aggregation number is the number of molecules per micelle
Ionic detergents are strong solubilizing agents that cause total membrane disruption by denaturing proteins by breaking protein-protein interactions and destroying protein activity and function. They are often used in biochemistry and molecular biology for a variety of applications, such as cell lysis and protein electrophoresis, and also for protein solubilization, disrupting protein-protein interactions, and membrane protein isolation. These types of detergents contain charged hydrophilic groups in their structure and are classified into cationic detergents (positively charged), anionic detergents (negatively charged), or zwitterionic detergents (both positive and negative changes) (see Figure 1).
| Sodium deoxycholate | SDS (sodium dodecyl sulfate), C12 | SDS (sodium dodecyl sulfate), Lauryl | |
| Properties | Anionic Useful for disrupting and dissociating protein interactions Frequently used in cell lysis buffers (RIPA buffer) | Anionic Strong lysis agent that works on a majority of cell types Not suitable for sensitive protein extraction due to its denaturing properties | |
| Popular for protein electrophoresis and solubilization Eliminate most all secondary chain lengths except C12 | Popular for denaturing polyacrylamide gel electrophoresis (SDS-PAGE) | ||
| Dialyzable | Yes | No | |
| Aggregation number* | 5 | 62 | |
| MW | 414.6 | 288.5 | |
| Micelle MW | 2,000 | 18,000 | |
| CMC % w/v (mM) | 0.083 to 0.249% (2-6) | 0.17-0.23% (6-8) | |
| Cloud point (°C) | Unknown | >100 | |
| Available size(s) | 25 g 5 g | 500 g | 100 g |
* Aggregation number is the number of molecules per micelle
Zwitterionic detergents can solubilize proteins without denaturing them, allowing proteins to maintain their native structure and function. They are used in biological and biochemical research, particularly for the extraction, solubilization, and stabilization of membrane proteins. These types of detergents are unique from ionic and non-ionic detergents, in that they contain both positively and negatively charged functional groups and have no net charge.
| CHAPS | CHAPSO | |
| Properties | Zwitterionic Mild lysis agent effective for the lysis of cultured mammalian cells Suitable for protecting the native state of proteins and maintaining protein activity | |
| Dialyzable | Yes | Yes |
| Aggregation number* | 10 | ~11 |
| MW | 615 | 630.9 |
| Micelle MW | 6,149 | Unknown |
| CMC % w/v (mM) | 0.49-0.62% (8-10) | 0.5% (8) |
| Cloud point (°C) | >100 | Unknown |
| Compatible with Detergent Removal Resin | Yes | No |
| Available size(s) | 5 g 100 g | 1 g 5 g |
* Aggregation number is the number of molecules per micelle
Selecting a detergent for protein solubilization
Selecting a detergent will be determined by their specific characteristics and the nature of the protein being studied. Detergents are classified according to their denaturing or non-denaturing properties. Denaturing detergents are designed to unfold and denature proteins, often used in applications such as protein purification and isolation. Non-denaturing detergents are formulated to maintain the native structure and function of proteins during solubilization, facilitating downstream applications that rely on the preserved state of the proteins.
Detergents are further classified into three categories: ionic (charged, either anionic or cationic), non-ionic (uncharged), or zwitterionic (having both positively and negatively charged groups but with a net charge of zero). In general, non-ionic and zwitterionic detergents are milder and less denaturing than ionic detergents and are used to solubilize membrane proteins where it is critical to maintain protein function and/or retain native protein-protein interactions for enzyme assays or immunoassays. CHAPS, a zwitterionic detergent, and the Triton-X series of non-ionic detergents are commonly used for these purposes. In contrast, ionic detergents are strong solubilizing agents and tend to denature proteins, thereby destroying protein activity and function (Figure 1).
Figure 1. Detergents used for cell lysis. Major characteristics of denaturing and non-denaturing detergents used for protein extraction.
The choice of detergent for cell lysis also depends on sample type. Animal cells, bacteria, and yeast all have different requirements for optimal lysis due to the presence or absence of a cell wall. Because of the dense and complex nature of animal tissues, they require both detergent and mechanical lysis. In addition to the choice of detergent, other important considerations for optimal cell lysis include the buffer, pH, salt concentration, and temperature. Consideration should be given to the compatibility of the chosen detergent with downstream applications. If the detergent used for lysis must be removed, then a dialyzable detergent should be selected.
Learn more: Detergents for Protein Extraction
Surfact-Amps Detergent Solutions
Surfact-Amps Detergent Solutions are easy-to-use 10% (w/v) solutions of highly purified detergents that can be utilized in routine and high-demand protein research methods and molecular biology techniques. These formulations provide high purity, quality, and stability. Unlike neat (undiluted) detergents, which are extremely viscous, Surfact-Amps 10% solutions are easy to pipet and can be accurately dispensed. The surfactant solutions are carefully prepared and packaged under nitrogen in glass ampules or non-leaching HDPE bottles, helping to ensure their stability and minimizing the accumulation of peroxides and degradation products.
Key features of protein solubilization detergents include:
- Accurate—precise 10% detergent solution in ultrapure water
- Easy-to-use—solution is simple to dispense and dilute for use
- Exceptionally pure—less than 1.0 µeq/mL peroxides and carbonyls
- Stable—packaged under inert nitrogen gas in glass ampules or HDPE bottles
Ordering information
See all: Cell lysis detergents
References
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Protein sample preparation technical handbook
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This modular, animated, and narrated eLearning course was developed to provide a succinct, contextual summary of common laboratory methods and techniques required to achieve optimal results in protein assays and experiments. There are knowledge checks throughout the course to test what you have learned.
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For Research Use Only. Not for use in diagnostic procedures.