In biochemical work, DL-Dithiothreitol is frequently used to protect biomolecules, denaturate proteins before acetone analysis (SDS-PAGE), and reduce disulfide bridges.

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- DTT acts as a strong reducing agent in vitro, specifically reducing disulfide bonds (S-S) in proteins and peptides to sulfhydryl groups (-SH). It is widely used to maintain proteins in a reduced, soluble state during experiments such as protein purification, electrophoresis, and enzyme assays [1]
- DTT prevents the formation of intramolecular or intermolecular disulfide bonds in proteins, which helps preserve protein structure and function in vitro experimental systems [1]

- In cell-related experiments, DTT is often added to cell lysis buffers to reduce disulfide bonds in cellular proteins, preventing protein aggregation and maintaining protein solubility for subsequent analyses (e.g., Western blot, protein quantification) [1]
- DTT can be used in cell culture systems at appropriate concentrations to maintain a reducing environment, though high concentrations may affect cell viability [1]

Absorption, Distribution and Excretion
Two male patients with advanced (uremic) infantile nephrotic cystinopathy (INC) received oral dithiothreitol (DTT) at a dose not exceeding 25 mg/kg body weight three times daily. Both patients underwent three consecutive observation periods: during thiol administration (8.5 months); during thiol discontinuation (8–9 months); and during thiol re-administration (7 months or longer)... Although chemical methods are not reliable in detecting and measuring DTT in biological fluids, preliminary evidence suggests that silanized derivatives of oxidized DTT can be detected in the urine of patients who received oral DTT. This finding indicates that thiols can be absorbed and excreted. Metabolism/Metabolites Two male patients with advanced (uremic) infantile nephrotic cystinopathy (INC) received oral dithiothreitol (DTT) at a dose not exceeding 25 mg/kg body weight three times daily. Both patients underwent three consecutive observation periods: thiol administration (8.5 months); thiol discontinuation (8-9 months); and thiol re-administration (7 months or longer)... Although chemical methods are not reliable for detecting and measuring DTT in biological fluids, preliminary evidence suggests that silanized derivatives of oxidized DTT can be detected in the urine of patients who took oral DTT. This finding indicates that thiols can be absorbed and excreted.

Toxicity Summary
Identification and Uses: 1,4-Dithiothreitol (DTT) is commonly used in biochemical experiments involving proteins or peptides to protect sulfhydryl groups from oxidation and reduce disulfide bonds between cysteine residues. It is also used to study disulfide exchange reactions of protein disulfide bonds, and DTT can keep glutathione in a reduced state. It has been used in experimental treatments for cystinosis or diseases caused by ionic or metal toxicity. Human Studies: DTT induces apoptosis in HL-60 cells. DTT is used to thin sputum in asthmatic patients. Two male patients with late-stage (uremia) infantile nephrotic cystinosis received oral DTT at doses not exceeding 25 mg/kg body weight three times daily. No other significant toxicities were observed except for nausea and vomiting within the maximum dose range. One subject died of uremia at month 24 of the study. Animal Studies: The inhibitory effects of dithiothreitol (DTT) on the heart and intestinal tissues of rats severely limit its application as an antioxidant in pharmacological studies to protect drugs susceptible to air oxidation. Dithiothreitol treatment can mimic the intracellular activation of Clostridium difficile's potent cytotoxin B.

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Interactions
Widely used thiol antioxidants (dithiothreitol, glutathione, and N-acetylcysteine) exhibit toxic activity against the human lymphocytic leukemia cell line HL60 when used in combination with hydroxycobalamin (vitamin B12). Following combined treatment with thiols and vitamin B12, early lysosomal instability and apoptosis occur. Caspase inhibitors can eliminate this cytotoxic effect. The iron chelator deferoxamine can partially prevent cell death, while lysosomal protease inhibitors and pepsin inhibitors offer no protection.
Arsenic is a naturally occurring toxic metalloid; drinking water containing As₂O₃ is believed to be associated with increased risk of neurotoxicity, liver damage, blackfoot disease, hypertension, and cancer. On the other hand, As₂O₃, as an ancient traditional Chinese medicine, has significant anticancer activity, particularly in the treatment of acute promyelocytic leukemia and in promoting chronic wound healing. However, the cytotoxicity of As₂O₃ against solid cancer cells (such as oral cancer cells) and its detailed mechanism of action remain largely unclear. In this study, we cultured four pairs of tumor and non-tumor cells from oral cancer patients and treated these cells with arsenic trioxide (As₂O₃) alone or in combination with dithiothreitol (DTT). The results showed that 0.5 μM As₂O₃ combined with 20 μM DTT significantly killed oral cancer cells but had no significant effect on non-tumor cells. Furthermore, As₂O₃ combined with DTT upregulated the expression of Bax and Bak, downregulated the expression of Bcl-2 and p53, and led to loss of mitochondrial membrane potential in oral cancer cells. On the other hand, As₂O₃ also induced endoplasmic reticulum stress and increased the expression levels of glucose regulatory protein 78, calpains 1, and 2. Our results indicate that DTT synergistically enhances the killing effect of As₂O₃ on oral cancer cells without toxicity to non-tumor cells. This combination therapy shows promise in the clinical treatment of oral cancer and warrants further investigation. Previous studies have found that vitamin B12b can significantly enhance the cytotoxic effect of ascorbic acid by catalyzing the generation of reactive oxygen species, while the antioxidant dithiothreitol (DTT), unlike catalase, cannot inhibit this cytotoxic effect. Therefore, this study investigated whether vitamin B12b could enhance the cytotoxic effect of DTT. The results showed that vitamin B12b significantly enhanced the cytotoxic effect of DTT. After the addition of vitamin B12b to DTT, the generation and accumulation of hydrogen peroxide in the culture medium rapidly reached a concentration of 260 μM within 7 minutes. The extracellular oxidative burst induced by the combined action of vitamin B12b and DTT (DTT + B12b) was accompanied by intracellular oxidative stress, lysosomal instability, and DNA damage. The accumulation of DNA damage led to the initiation of apoptosis, including the activation of caspase-3 and the release of cytochrome c. The antioxidants pyruvate and catalase completely inhibited DTT+vitamin B12b-induced oxidative stress and cell death. While the iron chelators deferoxamine and phenanthrene-1,000 did not reduce exogenous oxidative burst, they inhibited the genotoxic and cytotoxic effects of this combination, indicating that intracellular iron plays a crucial role in the cytotoxicity of this combination. Therefore, vitamin B12b significantly enhanced the cytotoxicity of DTT, catalyzing hydrogen peroxide production and inducing intracellular and extracellular oxidative stress, early lysosomal instability, and iron-dependent DNA damage. Inorganic trivalent arsenic compounds are ortho-thiol reactants, and dithiothreitol (DTT) is a well-known dithiol reactant. Interestingly, it has been reported that dithiothreitol (DTT) has both inhibitory and promoting effects on arsenic trioxide-induced apoptosis. The data we present now indicate that at high concentrations, DTT, dimercaptosuccinic acid (DMSA), and dimercaptopropanesulfonic acid (DMPS) all reduce arsenic trioxide-induced apoptosis in NB4 cells (a human promyelocytic leukemia cell line). Conversely, at low concentrations, DTT, DMSA, and DMPS increase arsenic trioxide-induced apoptosis. High concentrations (3 mM) of DTT reduce the growth-inhibiting effects of arsenic trioxide, methylarsenic acid (MMA(III)), and dimethylarsenic acid (DMA(III)) on NB4 cells, while low concentrations (0.1 mM) of DTT enhance this inhibitory effect. DMSA and DMPS are currently used as antidotes for acute arsenic poisoning. In experiments using human epithelial cell lines derived from arsenic target tissues such as the kidneys and bladder, these two dithiols have shown a reverse excitatory effect against arsenic toxicity in terms of DNA damage, micronucleus induction, apoptosis, and colony formation. The concentrations of these dithiols in the human body may be low after oral administration. Therefore, the current findings suggest that it is necessary to reassess the therapeutic effects of these dithiol compounds on arsenic poisoning. More complete data on 1,4-dithiothreitol (7 interactions) can be found on the HSDB record page. Non-human toxicity values: Mouse intramuscular LD50: 108 mg/kg Mouse intraperitoneal LD50: 154 mg/kg - Dithiothreitol (DTT) can be toxic through ingestion, inhalation, or skin absorption. The oral LD50 of DTT in rats is approximately 1500 mg/kg [1] - Exposure to DTT may irritate the eyes, skin, and respiratory tract. High concentrations can induce cellular oxidative stress [1]

[1]. Dithiothreitol, From Wikipedia.

1,4-Dithiothreitol is the threo-diastere of 1,4-dimercaptobutane-2,3-diol. It has the functions of a reducing agent, chelating agent and human metabolite. It is a dithiol and also a 1,4-dimercaptobutane-2,3-diol.
It has been reported that dithiothreitol exists in the human body and there is relevant data.
Dithiothreitol is a reagent commonly used in biochemical research. It can be used as a protective agent to prevent the oxidation of thiol (SH) and reduce disulfide bonds to dithiol.

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- Dithiothreitol is a small molecule thiol compound with the chemical formula C4H10O2S2 [1] - Its core mechanism depends on the reversible oxidation of two thiol groups to form disulfide bonds, which enables it to be used as a reducing agent in biochemical experiments [1] - Dithiothreitol is mainly a laboratory reagent rather than a therapeutic drug. It is commonly used in molecular biology, biochemistry and biotechnology research (e.g. SDS-PAGE, protein purification, antibody preparation) [1]
- DTT is more stable than other reducing agents (e.g. β-mercaptoethanol) and has a higher reducing power, thus it is suitable for experiments that require long-term maintenance of a reducing environment [1]

C4H10O2S2

154.2510

154.012

3483-12-3

DL-dithiothreitol-d6;850153-85-4;L-Dithiothreitol;16096-97-2;DL-dithiothreitol-d10;302912-05-6;DL-dithiothreitol-d10-1;203633-21-0

19001

White to off-white solid powder

1.3±0.1 g/cm3

364.5±42.0 °C at 760 mmHg

38-43ºC

174.2±27.9 °C

0.0±1.8 mmHg at 25°C

1.579

0.07

4

4

3

8

52

0

VHJLVAABSRFDPM-UHFFFAOYSA-N

InChI=1S/C4H10O2S2/c5-3(1-7)4(6)2-8/h3-8H,1-2H2

1,4-bis(sulfanyl)butane-2,3-diol

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.  (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ≥ 200 mg/mL (~1296.60 mM)
DMSO : ~100 mg/mL (~648.30 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (16.21 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (16.21 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (16.21 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 100 mg/mL (648.30 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)