You might be designing a cell lysis experiment to extract your target protein in its native form, or you have isolated cell membranes from which you want to solubilize a protein for further analyses. In these situations, detergents are your best buddies. Here are the details of twelve detergents that you can consider for your experimental needs. They are summarized just below in table 1.

We have listed a bunch of helpful detergents and their relevant properties – whether they are classified as zwitterionic/ nonionic/ cationic/ anionic, and if they are used for denaturing proteins or not. We also have their critical micelle concentration (CMC) expressed as weight/ unit volume (w/v) or molarity. For the nondenaturing detergents that are used to solubilize membranes, you would definitely want to use them at a concentration greater than the CMC so that it forms micelles to disrupt the lipid layers in the membranes.

There is also a really handy resource if you want a quick recap about the properties of detergents that help with their functions in experiments.

Table 1. Detergents and their attributes

Table 1. Detergents and their attributes
Detergent Classification Denaturing CMC
CHAPS Zwitterionic No 0.49% (w/v); 8-10 mM
Chlorhexidine Cationic Yes Not relevant in experiments
Chlorhexidine diacetate Cationic Yes 0.01 mM
(mostly not relevant in experiments)
Chlorhexidine HCl Cationic Yes Not relevant in experiments
Digitonin Non-ionic No 0.02-0.03% (w/v); < 0.5 mM
Dodecyl maltoside Non-ionic No 0.0087% (w/v) 0.17 mM
Deoxycholic acid Anionic Yes 4-8 mM;
Octylglucoside Non-ionic No 0.73% (w/v); 20-25 mM
Octylthioglucoside Non-ionic No 0.2772% (w/v); 4-9 mM
Glycochenodeoxycholic acid Anionic Yes Not relevant in experiments
1,2-Dipalmitoyl-racglycero-3-phosphocholine Cationic Yes 4.6 ± 0.5 x 10-10 M
Tridecyl beta-D-maltopyranoside Non-ionic No 0.033 mM; 0.0017% (w/v)



We have covered all you need to know about the basic concepts of how detergents work and why they are so important as reagents in bioscience experiments – in this article. Check it out if you want a detailed understanding of key parameters like critical micelle concentration (CMC), and how detergents are classified as anionic, cationic, zwitterionic or non-ionic.

This article is built around Table 1 shown above. We will briefly discuss the characteristics of these twelve detergents, which would be important for you when you are considering which detergents to use for your experiment.

 

CHAPS

CHAPS is the abbreviation of 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate, a zwitterionic detergent. Because it is non-denaturing, it is used in purifying proteins, especially membrane associated ones, in their native form.

If you use this detergent, you might be able to strike the perfect balance between being able to effectively lyse or permeabilize the cells and their membranes you have in your sample, while also preserving native structures of the proteins.

Uses: cell lysis and permeabilization while preserving native structure of sample proteins.

 

Chlorhexidine, chlorhexidine diacetate and chlorhexidine HCl

Chlorhexidine, chlorhexidine diacetate and chlorhexidine HCl are cationic detergents. Due to their positive charge, they bind to the negatively charged cell walls of bacteria, leading to their lysis. They are most effective in lysing cells of Gram-negative bacteria.

Oftentimes, lysing bacterial cells becomes necessary when you are using bacteria as cloning or expression hosts. And E. coli is the most used bacterium, which is Gram-negative.

Bacterial cells, especially Gram-negative bacteria, have a net negative charge on their cell surface due to negatively charged phosphoryl and carboxylate groups in their cell envelopes. The detergents chlorhexidine, chlorhexidine diacetate and chlorhexidine HCl, due to their positive charge, readily bind to these cell walls – and lyse them. Because of this reason, they can be a very handy reagent for lysing Gram-negative bacterial cells.

Uses: Lysing bacterial cells, particularly Gram-negative bacterial cells.

Figure 1. Cationic detergents (CD) with positive charge readily bind to the negatively charged cell-envelop of gram-negative bacteria, leading to membrane destabilization and lysis.

 

 

Deoxycholic acid

Deoxycholic acid is an anionic detergent mostly used in cell lysis as a component of lysis buffers because it disrupts cell and nuclear membranes. Naturally, it is a bile acid found in the intestine to emulsify dietary fats to aid in digestion.

 

Deoxycholic acid is a common component of cell lysis buffers such as RIPA buffer, for example. It helps to solubilize membrane proteins. In fact, deoxycholic acid’s role in purifying membrane proteins is leveraged by biotechnology industries. These industries use this detergent in the process of making outer membrane protein-based vaccines.

Uses: Cell lysis; solubilizing proteins from cell membranes.

 

Digitonin

Digitonin is a non-ionic detergent that can solubilize lipids. Because of this, digitonin is commonly used in cell lysis where the objective is to purify proteins or organelles in their native biological forms. Digitonin also permeabilizes cells without killing them, like in live cell labelling.

Detergents like digitonin form micelles which can dissolve lipids that are otherwise insoluble. And that helps a whole lot in breaking up the cell membranes composed of a phospholipid bilayer. I’d encourage you to look at this article, where we have schematically described the details of how detergent micelles help with membrane solubilization.

Uses: Lysing cells while not disrupting the native biological forms of proteins and organelles; cell permeabilization experiments.

1,2-Dipalmitoyl-rac-glycero-3-phosphocholine

1,2-Dipalmitoyl-rac-glycero-3-phosphocholine is cationic. However, unlike typical detergents it is seldom used for simple cell lysis or protein extraction experiments. Instead, it is used for creating liposomes or reconstituting bilayers composed of lipids and cholesterol.

Uses: Creating liposomes or reconstituting lipid and cholesterol bilayers.

 

Dodecyl maltoside

n-Dodecyl-β-D-maltoside (DDM) is a non-ionic detergent widely used for purifying proteins from membranes in their native forms. Chemically, this detergent has the sugar maltose connected via a glycoside bond.

The sugar head is hydrophilic while the alkyl tail is lipophilic – just like how biological membranes are. This chemical property of having both hydrophobic as well as hydrophilic moieties makes it ideal to break up membranes.

Consider an experiment where you want to purify a certain membrane-associated protein to see which other proteins are bound to it. For this purpose, you’d need to purify the protein in its native form because if it is denatured, then all protein-protein interactions are destroyed.

In such a situation, DDM would be very helpful. You’d first precipitate the membrane fraction after cell lysis. And then, you’d need to resuspend that precipitate in a solution of a mild non-ionic detergent like DDM, which would solubilize proteins out of the membrane fraction while still preserving the protein-protein interactions.

Uses: Solubilizing proteins from cell membranes in their native form.

 

Glycochenodeoxycholic acid

Glycochenodeoxycholic acid is a bile acid that is used in experiments as an anionic detergent for cell lysis and to solubilize lipids for disrupting membranes. It is also used in some cell cultures involving cells of the digestive system such as hepatocytes.

Bile acids, including Glycochenodeoxycholic acid, help break down fats and aid in intestinal digestion. This property is utilized in lab experiments where they are used to dissolve lipids in cell membrane, lysing the cells in the process.

Uses: Cell lysis in denaturing conditions. Also used in some special cell cultures’ media.

 

Octylglucoside and Octylthioglucoside

Octylglucoside and octylthioglucoside are non-ionic detergents used for purifying membrane proteins in non-denaturing conditions. Chemically, they have a glycosidic bond between glucose and octanol.

The sugar part of these detergents serves as the hydrophilic portion, while the long acetyl chain composed of eight carbon atoms is the hydrophobic portion. So, if you see, the structures of these compounds have a stark resemblance to the hydrophobic-hydrophilic dual nature of membranes. This similarity is structure helps these detergents get incorporated into the membranes and destabilize them – ultimately lysing them partially or completely.

Uses: Extracting and purifying cell membrane proteins without disrupting protein-protein interactions and preserving native biological structures of the extracted proteins.

 

Tridecyl beta-D-maltopyranoside

Tridecyl beta-D-maltopyranoside is a non-ionic detergent that has a glycosidic linkage involving the sugar maltose. It is primarily used for membrane protein experiments that require mild, non-denaturing conditions.

Uses: membrane protein extraction and crystallography, micellar electrokinetic chromatography (used in pharmaceutical chemistry).

 

In this article, we described 12 detergents that Goldbio offers for your experiments. This is an extension of what we described in this preceding article about the modes of actions of different detergents, their classification and so on. So now you know not only how detergents might be helpful in your experiments in general, but also how specific detergents can help with your experimental needs.

References

Stetsenko and Guskov. 2017. Crystals. An Overview of the Top Ten Detergents Used for Membrane Protein Crystallization. 7(7), 197; https://doi.org/10.3390/cryst7070197

Wycisk and Urner. 2023. Protocol to test the utility of detergents for E. coli membrane protein extraction and delipidation. Protocol. doi: 10.1016/j.xpro.2023.102146

Tao et al. 2018. Purifying natively folded proteins from inclusion bodies using sarkosyl, Triton X-100, and CHAPS. Biotechniques. 48. https://doi.org/10.2144/000113304