07 Detergents

Detergent Screening Set (first choice-II)

Detergent

  • Product code
    DS06  Detergent Screening Set (first choice-II)
Unit size Price Item Code
1 set DS06-10
Component
1 set CHAPS,
n-Dodecyl-β-D-maltoside,
n-Octyl-β-D-glucoside,
Sodium cholate(purifi
200 mg x 1
200 mg x 1
200 mg x 1
200 mg x 1
200 mg x 1

Product Description

Detergent Screening Sets are used to find the best detergent for an application, such as enzyme stabilization, specific membrane protein isolation, assay development, protein crystallization, protein refolding, and so on. Detergent Screening Set (first choice-II) contains commonly used detergents for membrane protein isolation. Detergent Screening Set (for crystallization) contains detergents commonly used for protein crystallization.

Introduction

The phospholipid bilayer is the basic structure of the cell membrane. The most important functions of cells include transportation of substances, energy exchange, and transmission of information. These functions are conducted at the cell membrane by membrane proteins. In membrane biochemistry research, membrane proteins are solubilized and purified to study their structure and function. Proteins bound to cell membranes have hydrophobic sites buried within the phospholipid bilayers and hydrophilic sites facing toward the water layer. Detergents are used to isolate large insoluble molecules such as proteins. Detergents interact with the hydrophobic sites of proteins, which are then solubilized in the water layer, thus separating membrane proteins. It is important to choose a detergent that does not disrupt the bioactivities of target proteins. A detergent requires the following characteristics to be suitable for isolation of

 

membrane proteins:
1. Sufficient protein solubilization capability
2. No denaturing or inactivation of proteins
3. No interference with protein activities
4. No precipitation at 4ºC
5. Appropriate critical micelle concentrations (CMC) and micelle size
6. No absorption in the UV region
7. No toxicity
8. Availability of detergent detection methods
9. Non-ionic detergent if ion exchange chromatography is used

 

In the past, polyoxyethylene ether non-ionic detergents were widely used. These detergents, however, had several problems, such as denaturation of proteins and low CMC value, which cannot be separated easily by dialysis. n-Octyl-β-D-glucoside, n-Octyl-β-Dthioglucoside, CHAPS, and CHAPSO eliminate these problems and are widely used today. Most of the current detergents are non-ionic and easily applied to ion exchange chromatography purification. deoxy-BIGCHAP is a non-ionic detergent possessing deoxycholic acid and a gluconamide polar group. It has a high CMC value of 1.4 mM and can be easily separated by dialysis. Because its UV absorbance is low, it can be used for the determination of proteins. deoxy-BIGCHAP has been used for the extraction of opioid receptors from neuroblastoma or hybrid cells of glyoma. It has also been applied to adenylate cyclase or acetyltransferase. These detergents are also widely used to solubilize chromophores or to stabilize enzymes in diagnostic analyses and biochemical assays.

Trials of various kinds of detergents are needed to find the appropriate detergent for each study. Dojindo’s Detergent Screening Sets, which contain assorted packages of detergents, are available for use in these trials.

Technical info

Detergents are amphipathic compounds, with both lipophobic and lipophilic sites, that will form micelles above a critical concentration that is specific to each detergent. This is called the critical micelle concentration (CMC). The solubilizing abilities of detergents increase dramatically above their CMC values. After extracting membrane proteins, detergents can be easily removed by dilution, and then dialysis.

Table 1 Molecular Weight and Critical Micelle Concentration of Detergents

 

Specification

n-Dodecyl-β-D-maltoside:    To pass the test

n-Octyl-β-D-glucoside:    To pass the test

Sodium cholate(purified):    To pass the test

MEGA-8:    To pass the test

References

Open References

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13) K. A. Williams, U. Geldmacher-Kaufer, E. Padan, S. Schuldiner and W. Kuhlbrandt, "Projection Structure of NhaA, a Secondary Transporter from Escherichia Coli, at 4.0Å Resolution", EMBO J., 1999, 18, 3558.
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Handling and storage condition

Handling and storage condition
0-5°C, Protect from moisture
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Product Classification

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