DTNB

SKU: D0156

Category: Non Fluorescent Detection & Chromogene Detection

Synonyms: Ellman's Reagent , 5,5'-Dithiobis(2-nitrobenzoic acid) , 3-Carboxy-4-nitrophenyl disulfide , 6,6'-Dinitro-3,3'-dithiodibenzoic acid , Bis(3-carboxy-4-nitrophenyl) disulfide; 69-78-3
CAS-Number: C₁₄H₈N₂O₈S₂
Molecular Formula: 396.35
Molecular Weight

Product Data
Handling
Safety Information
Details

Specifications

Purity: ≥98% (Titration)
Identity: 1H-NMR
Appearance: Yellow powder

Properties

Melting Point: 240-245 °C
Solvents: ethanol, dioxane, phosphate buffer

Handling Advice: Protect from light and moisture.
Use / Stability: Stable for at least 2 years after receipt when stored at RT.
Long Term Storage: RT
Short Term Storage: RT
Shipping: AMBIENT

Transportation: Packing Group III
Signal word: Warning
GHS Symbol: GHS07
Precautionary statements: P261-P305 + P351 + P338
Hazard statements: H315-H319-H335

Description: A sensitive reagent for measuring the free sulfhydryl content in proteins, peptides, and tissues. Used to characterize reactive thiol groups and photometric determination of thiols and for measuring low-molecular mass thiols such as glutathione in both pure solutions and biological samples, such as blood. It can also measure the number of thiol groups on proteins. Through reaction with aliphatic thiol groups a mixed disulfide of protein thiol and one mole of 2-nitro-5-thiobenzoate per mole of protein sulfhydryl group is being formed. DTNB has little absorbance. Reaction with -SH groups on proteins (from any solvent accessible Cys) under mild alkaline conditions (pH 7-8) produces the 2-nitro-5-thiobenzoate anion, which gives an intense yellow color with an absorption maximum at 409.5nm (Extinction coefficient: 14150 M-1*cm-1). Sensitive to various buffer ions, therefore, the extinction coefficient used to calculate the number of sulfhydryl groups must be matched to the reaction conditions. In case the thiol groups are in disulfide bonds, they must be reduced under anaerobic conditions prior to reaction with DTNB.
InChi: InChI=1S/C14H8N2O8S2/c17-13(18)9-5-7(1-3-11(9)15(21)22)25-26-8-2-4-12(16(23)24)10(6-8)14(19)20/h1-6H,(H,17,18)(H,19,20)
References: (1) G.L. Ellman: Arch. Biochem. Biophys. 82, 70 (1959) , (2) H.B. Collier: Anal. Biochem. 56, 310 (1973) , (3) P.W. Riddles, et al.: Meth. Enzymol. 91, 49 (1983) , (4) C.K. Riener, et al.: Anal. Bioanal. Chem. 373, 266 (2002) , (5) J. Sedlak & R.H. Lindsay: Anal. Biochem. 25, 192 (1968) , (6) G.L. Ellman: Biochem. Pharmacol. 7, 88 (1961) , (7) M.J. Gething & B.E. Davidson: Eur. J. Biochem. 30, 352 (1972) , (8) A.F.S.A. Habeeb: Meth. Enzymol. 25, 457 (1972) , (9) D. Ming, et al.: Biotechniques 18, 808 (1995)
InChi Key: KIUMMUBSPKGMOY-UHFFFAOYSA-N
Smiles: O=[N+]([O-])C1=CC=C(SSC2=CC=C([N+]([O-])=O)C(C(O)=O)=C2)C=C1C(O)=O

A sensitive reagent for measuring the free sulfhydryl content in proteins, peptides, and tissues. Used to characterize reactive thiol groups and photometric determination of thiols and for measuring low-molecular mass thiols such as glutathione in both pure solutions and biological samples, such as blood. It can also measure the number of thiol groups on proteins. Through reaction with aliphatic thiol groups a mixed disulfide of protein thiol and one mole of 2-nitro-5-thiobenzoate per mole of protein sulfhydryl group is being formed. DTNB has little absorbance. Reaction with -SH groups on proteins (from any solvent accessible Cys) under mild alkaline conditions (pH 7-8) produces the 2-nitro-5-thiobenzoate anion, which gives an intense yellow color with an absorption maximum at 409.5nm (Extinction coefficient: 14150 M-1*cm-1). Sensitive to various buffer ions, therefore, the extinction coefficient used to calculate the number of sulfhydryl groups must be matched to the reaction conditions. In case the thiol groups are in disulfide bonds, they must be reduced under anaerobic conditions prior to reaction with DTNB.

SKU: D0156 Category: Non Fluorescent Detection & Chromogene Detection

Additional information

Additional information

Synonyms	Ellman's Reagent, 5,5'-Dithiobis(2-nitrobenzoic acid), 3-Carboxy-4-nitrophenyl disulfide, 6,6'-Dinitro-3,3'-dithiodibenzoic acid, Bis(3-carboxy-4-nitrophenyl) disulfide
Purity	≥98% (Titration)
Appearance	Yellow powder
CAS-Number	69-78-3
Molecular Formula	C14H8N2O8S2
Molecular Weight	396.35
Identity	1H-NMR
Solvents	ethanol, dioxane, phosphate buffer
Melting Point	240-245 °C
Smiles	O=[N+]([O-])C1=CC=C(SSC2=CC=C([N+]([O-])=O)C(C(O)=O)=C2)C=C1C(O)=O
InChi Key	KIUMMUBSPKGMOY-UHFFFAOYSA-N
Shipping	AMBIENT
Short Term Storage	RT
Long Term Storage	RT
Handling Advice	Protect from light and moisture.
Use / Stability	Stable for at least 2 years after receipt when stored at RT.
Hazard statements	H315-H319-H335
Precautionary statements	P261-P305 + P351 + P338
GHS Symbol	GHS07
Signal word	Warning
Transportation	Packing Group III
References	(1) G.L. Ellman: Arch. Biochem. Biophys. 82, 70 (1959), (2) H.B. Collier: Anal. Biochem. 56, 310 (1973), (3) P.W. Riddles, et al.: Meth. Enzymol. 91, 49 (1983), (4) C.K. Riener, et al.: Anal. Bioanal. Chem. 373, 266 (2002), (5) J. Sedlak & R.H. Lindsay: Anal. Biochem. 25, 192 (1968), (6) G.L. Ellman: Biochem. Pharmacol. 7, 88 (1961), (7) M.J. Gething & B.E. Davidson: Eur. J. Biochem. 30, 352 (1972), (8) A.F.S.A. Habeeb: Meth. Enzymol. 25, 457 (1972), (9) D. Ming, et al.: Biotechniques 18, 808 (1995)
InChi	InChI=1S/C14H8N2O8S2/c17-13(18)9-5-7(1-3-11(9)15(21)22)25-26-8-2-4-12(16(23)24)10(6-8)14(19)20/h1-6H,(H,17,18)(H,19,20)
Quantity	1 g, 5 g, 25 g, Bulk