Synonym
DTNB; Ellman's Reagent; Dithiobisnitrobenzoic acid;
IUPAC/Chemical Name
5,5'-Dithiobis(2-nitrobenzoic acid)
InChi Key
KIUMMUBSPKGMOY-UHFFFAOYSA-N
InChi Code
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)
SMILES Code
O=C(O)C1=CC(SSC2=CC=C([N+]([O-])=O)C(C(O)=O)=C2)=CC=C1[N+]([O-])=O
Purity
>98% (or refer to the Certificate of Analysis)
Shipping Condition
Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.
Storage Condition
Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).
Solubility
Soluble in DMSO
Shelf Life
>3 years if stored properly
Drug Formulation
This drug may be formulated in DMSO
Stock Solution Storage
0 - 4 C for short term (days to weeks), or -20 C for long term (months).
HS Tariff Code
2934.99.03.00
Biological target:
DTNB (Ellman’s Reagent) is a chemical used to quantify the number or concentration of thiol groups.
In vitro activity:
The enzyme had an NH2-terminal sequence of TQSNPPSWGLDRIDQTTAFTKACSIKY, thus sharing high homology with those of Streptomyces proteases. The results showed that this protease was completely inhibited by phenylmethanesulfonyl fluoride (PMSF), diiodopropyl fluorophosphates (DFP), and partially inhibited by 5,5-dithio-bis-(2-nitro benzoic acid) (DTNB), which strongly suggested its belonging to the serine thiol protease family.
Reference: Int J Biol Macromol. 2015 Aug;79:871-82. https://pubmed.ncbi.nlm.nih.gov/26056991/
In vivo activity:
The effects of DTNB on changes in intracellular Ca(2+) content in slices of rat cortex induced by 2- or 10-min anoxia were studied fluorometrically. DTNB (200 microM) prevented excessive intracellular Ca(2+) accumulation during and after 10-min anoxia and moderate increase in Ca(2+) content induced by 2-min anoxia.
Reference: Bull Exp Biol Med. 2001 Jun;131(6):526-8. https://pubmed.ncbi.nlm.nih.gov/11586396/
|
Solvent |
mg/mL |
mM |
| Solubility |
| DMSO |
89.5 |
225.81 |
| Ethanol |
79.0 |
199.32 |
Note: There can be variations in solubility for the same chemical from different vendors or different batches from the same vendor. The following factors can affect the solubility of the same chemical: solvent used for crystallization, residual solvent content, polymorphism, salt versus free form, degree of hydration, solvent temperature. Please use the solubility data as a reference only. Warming and sonication will facilitate dissolving. Still have questions? Please contact our Technical Support scientists.
Preparing Stock Solutions
The following data is based on the
product
molecular weight
396.34
Batch specific molecular weights may vary
from batch to batch
due to the degree of hydration, which will
affect the solvent
volumes required to prepare stock solutions.
| Concentration / Solvent Volume / Mass |
1 mg |
5 mg |
10 mg |
| 1 mM |
1.15 mL |
5.76 mL |
11.51 mL |
| 5 mM |
0.23 mL |
1.15 mL |
2.3 mL |
| 10 mM |
0.12 mL |
0.58 mL |
1.15 mL |
| 50 mM |
0.02 mL |
0.12 mL |
0.23 mL |
Formulation protocol:
1. Ben Elhoul M, Zaraî Jaouadi N, Rekik H, Bejar W, Boulkour Touioui S, Hmidi M, Badis A, Bejar S, Jaouadi B. A novel detergent-stable solvent-tolerant serine thiol alkaline protease from Streptomyces koyangensis TN650. Int J Biol Macromol. 2015 Aug;79:871-82. doi: 10.1016/j.ijbiomac.2015.06.006. Epub 2015 Jun 6. PMID: 26056991.
2. Liu JP, Cen SY, Xue Z, Wang TX, Gao Y, Zheng J, Zhang C, Hu J, Nie S, Xiong Y, Guan KL, Yuan HX. A Class of Disulfide Compounds Suppresses Ferroptosis by Stabilizing GPX4. ACS Chem Biol. 2022 Dec 16;17(12):3389-3406. doi: 10.1021/acschembio.2c00445. Epub 2022 Nov 29. PMID: 36446024.
3. Semenov DG, Samoilov MO, Lazarevich EV. DTNB inhibits calcium response of rat brain cortical slices to anoxia of various duration. Bull Exp Biol Med. 2001 Jun;131(6):526-8. doi: 10.1023/a:1012334031357. PMID: 11586396.
4. Diez N, Barber A, Ponz F. Effect of DTNB on rat intestinal galactose transport in vivo. Rev Esp Fisiol. 1991 Jun;47(2):69-73. PMID: 1924965.
In vitro protocol:
1. Ben Elhoul M, Zaraî Jaouadi N, Rekik H, Bejar W, Boulkour Touioui S, Hmidi M, Badis A, Bejar S, Jaouadi B. A novel detergent-stable solvent-tolerant serine thiol alkaline protease from Streptomyces koyangensis TN650. Int J Biol Macromol. 2015 Aug;79:871-82. doi: 10.1016/j.ijbiomac.2015.06.006. Epub 2015 Jun 6. PMID: 26056991.
2. Liu JP, Cen SY, Xue Z, Wang TX, Gao Y, Zheng J, Zhang C, Hu J, Nie S, Xiong Y, Guan KL, Yuan HX. A Class of Disulfide Compounds Suppresses Ferroptosis by Stabilizing GPX4. ACS Chem Biol. 2022 Dec 16;17(12):3389-3406. doi: 10.1021/acschembio.2c00445. Epub 2022 Nov 29. PMID: 36446024.
In vivo protocol:
1. Semenov DG, Samoilov MO, Lazarevich EV. DTNB inhibits calcium response of rat brain cortical slices to anoxia of various duration. Bull Exp Biol Med. 2001 Jun;131(6):526-8. doi: 10.1023/a:1012334031357. PMID: 11586396.
2. Diez N, Barber A, Ponz F. Effect of DTNB on rat intestinal galactose transport in vivo. Rev Esp Fisiol. 1991 Jun;47(2):69-73. PMID: 1924965.
1: Zacharis CK, Tzanavaras PD. Liquid chromatography coupled to on-line post column derivatization for the determination of organic compounds: a review on instrumentation and chemistries. Anal Chim Acta. 2013 Oct 10;798:1-24. doi: 10.1016/j.aca.2013.07.032. Epub 2013 Jul 16. Review. PubMed PMID: 24070479.
2: Nagy P, Pálinkás Z, Nagy A, Budai B, Tóth I, Vasas A. Chemical aspects of hydrogen sulfide measurements in physiological samples. Biochim Biophys Acta. 2014 Feb;1840(2):876-91. doi: 10.1016/j.bbagen.2013.05.037. Epub 2013 Jun 14. Review. PubMed PMID: 23769856.
3: Turell L, Radi R, Alvarez B. The thiol pool in human plasma: the central contribution of albumin to redox processes. Free Radic Biol Med. 2013 Dec;65:244-53. doi: 10.1016/j.freeradbiomed.2013.05.050. Epub 2013 Jun 7. Review. PubMed PMID: 23747983; PubMed Central PMCID: PMC3909715.
4: Winther JR, Thorpe C. Quantification of thiols and disulfides. Biochim Biophys Acta. 2014 Feb;1840(2):838-46. doi: 10.1016/j.bbagen.2013.03.031. Epub 2013 Apr 6. Review. PubMed PMID: 23567800; PubMed Central PMCID: PMC3766385.
5: Liu Y, Li Y, Yu S, Zhao G. Recent advances in the development of thioredoxin reductase inhibitors as anticancer agents. Curr Drug Targets. 2012 Oct;13(11):1432-44. Review. PubMed PMID: 22876886.
6: Laragione T, Gianazza E, Tonelli R, Bigini P, Mennini T, Casoni F, Massignan T, Bonetto V, Ghezzi P. Regulation of redox-sensitive exofacial protein thiols in CHO cells. Biol Chem. 2006 Oct-Nov;387(10-11):1371-6. Review. PubMed PMID: 17081109.
7: Stetler-Stevenson WG, Talano JA, Gallagher ME, Krutzsch HC, Liotta LA. Inhibition of human type IV collagenase by a highly conserved peptide sequence derived from its prosegment. Am J Med Sci. 1991 Sep;302(3):163-70. Review. PubMed PMID: 1656751.
