Dansyl chloride: Difference between revisions
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In addition, these adducts are, in general, good acceptors for [[fluorescence resonance energy transfer]] from the amino acid [[tryptophan]], allowing this labeling technique to be used in investigating protein flexibility. The fluorescence of these sulfonamide adducts can be enhanced by adding cycloheptaamylose.<ref>{{cite journal |author=Kinoshita T, Iinuma F, Tsuji A |title=Microanalysis of proteins and peptides. I. Enhancement of the fluorescence intensity of dansyl amino acids and dansyl proteins in aqueous media and its application to assay of amino acids and proteins |journal=Chem. Pharm. Bull. |volume=22 |issue=10 |pages=2413–20 |year=1974 |pmid=4468087}}</ref> It is important to note that dansyl chloride is unstable in [[dimethyl sulfoxide]], which should never be used to prepare solutions of the reagent. |
In addition, these adducts are, in general, good acceptors for [[fluorescence resonance energy transfer]] from the amino acid [[tryptophan]], allowing this labeling technique to be used in investigating protein flexibility. The fluorescence of these sulfonamide adducts can be enhanced by adding cycloheptaamylose.<ref>{{cite journal |author=Kinoshita T, Iinuma F, Tsuji A |title=Microanalysis of proteins and peptides. I. Enhancement of the fluorescence intensity of dansyl amino acids and dansyl proteins in aqueous media and its application to assay of amino acids and proteins |journal=Chem. Pharm. Bull. |volume=22 |issue=10 |pages=2413–20 |year=1974 |pmid=4468087}}</ref> It is important to note that dansyl chloride is unstable in [[dimethyl sulfoxide]], which should never be used to prepare solutions of the reagent. |
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The extinction coefficient of dansyl derivatives are important for measuring their concentration in solution. A variety of extinction coefficient values have been used. Some of the values are used to estimate the degree of protein labeling that was achieved. Other values may be used to determine a precise concentration of a stock solution. For all of the studies below, the absorption value is always taken at the ''maximum'' that appears between 320 nm and 340 nm. The peak is broad, but trivial errors in wavelength calibration can be avoided simply by taking the value at the maximum instead of strictly at 330 nm. |
The extinction coefficient of dansyl derivatives are important for measuring their concentration in solution. A variety of extinction coefficient values have been used. Some of the values are used to estimate the degree of protein labeling that was achieved. Other values may be used to determine a precise concentration of a stock solution. See the table below for specific values and their uses. |
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For all of the studies below, the absorption value is always taken at the ''maximum'' that appears between 320 nm and 340 nm. The peak is broad, but trivial errors in wavelength calibration can be avoided simply by taking the value at the maximum instead of strictly at 330 nm. |
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| title = The active center of chymotrypsin: 1. Labelling with a fluorescent dye |
| title = The active center of chymotrypsin: 1. Labelling with a fluorescent dye |
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| journal = Biochimica et biophysica acta |
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| date = 1956}} |
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|Used to determine degree of labeling in chymotrypsin |
| Use for DNSC-protein conjugates; Used to determine degree of labeling in chymotrypsin and ovalbumin |
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| title = Dansyl labeled proteins |
| title = Dansyl labeled proteins |
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| journal = Analytical Biochemistry |
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| date = 1968 }} |
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| In bicarbonate buffer; maximum is shifted to ~315 nm |
| In bicarbonate buffer; maximum is shifted to ~315 nm |
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| title = Molecular Probes Handbook: Coumarins, Pyrenes and Other Ultraviolet Light–Excitable Fluorophores (Section 1.7) |
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| url = http://www.invitrogen.com/site/us/en/home/References/Molecular-Probes-The-Handbook/Fluorophores-and-Their-Amine-Reactive-Derivatives/Coumarins-Pyrenes-and-Other-Ultraviolet-Light-Excitable-Fluorophores.html |
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Revision as of 16:13, 12 June 2009
| |
| Names | |
|---|---|
| IUPAC name
5-(dimethylamino)naphthalene-1-sulfonyl chloride
| |
| Identifiers | |
| ECHA InfoCard | 100.009.175 |
| RTECS number |
|
CompTox Dashboard (EPA)
|
|
| Properties | |
| C12H12ClNO2S | |
| Molar mass | 269.75 g/mol |
| Melting point | 70 °C (158 °F; 343 K) |
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
| |
Dansyl chloride or 5-(dimethylamino)naphthalene-1-sulfonyl chloride is a reagent that reacts with primary amino groups in both aliphatic and aromatic amines to produce stable blue- or blue-green–fluorescent sulfonamide adducts. Dansyl chloride is widely used in protein sequencing and amino acid analysis.[2][3]
Dansyl chloride may also be denoted DNSC. Likewise, a similar derivative, dansyl amide is known as DNSA.
In addition, these adducts are, in general, good acceptors for fluorescence resonance energy transfer from the amino acid tryptophan, allowing this labeling technique to be used in investigating protein flexibility. The fluorescence of these sulfonamide adducts can be enhanced by adding cycloheptaamylose.[4] It is important to note that dansyl chloride is unstable in dimethyl sulfoxide, which should never be used to prepare solutions of the reagent.
The extinction coefficient of dansyl derivatives are important for measuring their concentration in solution. A variety of extinction coefficient values have been used. Some of the values are used to estimate the degree of protein labeling that was achieved. Other values may be used to determine a precise concentration of a stock solution. See the table below for specific values and their uses.
For all of the studies below, the absorption value is always taken at the maximum that appears between 320 nm and 340 nm. The peak is broad, but trivial errors in wavelength calibration can be avoided simply by taking the value at the maximum instead of strictly at 330 nm.
| Species | Extinction Coefficient [1/M * 1/cm] | Reference | Notes |
|---|---|---|---|
| DNSC-protein | 3300 | Hartley, BS (1956). "The active center of chymotrypsin: 1. Labelling with a fluorescent dye". Biochimica et biophysica acta. 21: 58–70. {{cite journal}}: Unknown parameter |coauthors= ignored (|author= suggested) (help)
|
Use for DNSC-protein conjugates; Used to determine degree of labeling in chymotrypsin and ovalbumin |
| DNSC | 4350 | Chen, RF (1968). "Dansyl labeled proteins". Analytical Biochemistry. 25: 412–416. | In bicarbonate buffer; maximum is shifted to ~315 nm |
| DNSC | 4000 | Molecular Probes Handbook: Coumarins, Pyrenes and Other Ultraviolet Light–Excitable Fluorophores (Section 1.7), retrieved 12 June 2009 | Conditions are not given; no reference to the source of this value |
References
- ^ "MSDS". Sigma-Aldrich. Retrieved 2007-12-02.
- ^ Walker JM (1994). "The dansyl method for identifying N-terminal amino acids". Methods Mol. Biol. 32: 321–8. doi:10.1385/0-89603-268-X:321. PMID 7951732.
- ^ Walker JM (1994). "The Dansyl-Edman method for peptide sequencing". Methods Mol. Biol. 32: 329–34. doi:10.1385/0-89603-268-X:329. PMID 7951733.
- ^ Kinoshita T, Iinuma F, Tsuji A (1974). "Microanalysis of proteins and peptides. I. Enhancement of the fluorescence intensity of dansyl amino acids and dansyl proteins in aqueous media and its application to assay of amino acids and proteins". Chem. Pharm. Bull. 22 (10): 2413–20. PMID 4468087.
{{cite journal}}: CS1 maint: multiple names: authors list (link)
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