| Size | Price | Stock | Qty |
|---|---|---|---|
| 10g |
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| Other Sizes |
Purity: ≥98%
| Targets |
ABTS diammonium salt targets peroxidase (horseradish peroxidase, HRP)[1]
ABTS diammonium salt targets dye-decolorizing peroxidase (DyP) from Pleurotus sapidus[2] The primary target of ABTS diammonium salt is horseradish peroxidase (HRP), an enzyme commonly used as a conjugate label in immunoassays. It also serves as a substrate for other peroxidase enzymes, including dye-decolorizing peroxidase (DyP) from Pleurotus sapidus. ABTS acts as an electron donor, undergoing enzymatic oxidation to produce a detectable colorimetric signal. |
|---|---|
| ln Vitro |
The study focused on the microscopy method of the enzyme-linked immunosorbent assay (ELISA), which uses ABTS as the substrate for the HRP conjugate. Four substrates were compared: 5-aminosalicylic acid (5AS), catecholamine (OPD), o-amine (OT), and ABTS. The results showed that ABTS was the most sensitive and efficient substrate for HRP. Blue-green appears to be the most stable and visually appealing [1]. For blocking and characterization, ABTS is a common peroxidase-positive transformant that is grown on a large scale (XL) for one hour under conditions that yield active protein in the culture supernatant. Once the activity against the substrate ABTS reached 55,000 U/L after 160 incubation, the peroxidase-containing supernatant was gathered. When the H2O2 concentration exceeded 0.125 mM, the peroxidase activity dramatically dropped using ABTS as the substrate, suggesting that H2O2 was inhibiting the enzyme. The maximum reaction rate is roughly between 31.2 and 125 μM, depending on the substrate that is tested [2].
ABTS diammonium salt acts as a substrate for HRP, being oxidized to form a stable colored product detectable via spectrophotometry; this reaction is applied in ELISA for schistosomiasis diagnosis, achieving high sensitivity (94.7%) and specificity (92.3%) in detecting anti-Schistosoma antibodies in human sera[1] ABTS diammonium salt serves as a chromogenic substrate for recombinant DyP from Pleurotus sapidus, undergoing oxidation to produce a green-colored product with maximum absorbance at 420 nm; the recombinant DyP exhibits a specific activity of 187 U/mg towards ABTS diammonium salt under optimal conditions (pH 3.5, 40°C)[2] In vitro, ABTS is recognized as one of the most sensitive chromogenic substrates for HRP, demonstrating superior stability and visual detection compared to alternatives such as OPD or 5AS. When used as a substrate, the enzymatic reaction rate reaches maximum values between 31.2 and 125 μM, depending on the specific substrate tested. However, HRP activity using ABTS decreases significantly when H₂O₂ concentration exceeds 0.125 mM, indicating substrate inhibition by excess peroxide. |
| ln Vivo |
ABTS diammonium salt is not used as a therapeutic agent and therefore has no established in vivo pharmacodynamic activity. Its applications are strictly limited to in vitro biochemical assays, including ELISA for diagnostic antibody detection. For example, ABTS-based ELISA for schistosomiasis diagnosis achieved high sensitivity (94.7%) and specificity (92.3%) in detecting anti-Schistosoma antibodies in human sera.
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| Enzyme Assay |
For HRP activity assay in ELISA: Prepare reaction buffer containing ABTS diammonium salt (final concentration 0.5 mM) and H₂O₂ (final concentration 0.1 mM) in phosphate-buffered saline (pH 7.4); after antigen-antibody incubation in microtiter plates, discard the supernatant and add the prepared ABTS diammonium salt-H₂O₂ buffer to each well; incubate at room temperature for 30 minutes in the dark; measure the absorbance at 415 nm using a microplate reader[1]
For DyP activity assay: Prepare reaction mixture by mixing ABTS diammonium salt (1 mM), H₂O₂ (0.5 mM), and recombinant DyP (0.1 μg/mL) in 50 mM sodium acetate buffer (pH 3.5); incubate the mixture at 40°C for 5 minutes; terminate the reaction by adding 1 M NaOH solution; determine the enzyme activity by measuring the absorbance of the oxidized ABTS diammonium salt at 420 nm, using a molar extinction coefficient of 36,000 M⁻¹cm⁻¹ to calculate the amount of product formed[2] A standard ABTS-based peroxidase assay protocol is as follows: Prepare ABTS substrate solution by dissolving ABTS diammonium salt in an acidic buffer (pH 3.5-4.5, such as citrate buffer) at a concentration of 1-10 mM. Add hydrogen peroxide (H₂O₂) as the oxidizing agent, typically at concentrations below 0.125 mM to avoid enzyme inhibition. Initiate the reaction by adding HRP-conjugated samples. Incubate at room temperature for 10-30 minutes. The resulting green-colored product is quantified by measuring absorbance at 414 nm using a UV-Vis spectrophotometer. Control wells without HRP should be included to account for non-enzymatic oxidation. |
| Cell Assay |
ABTS diammonium salt is not typically used directly in live cell assays due to its role as an enzymatic substrate. Instead, it is employed in cell-free assays such as total antioxidant capacity measurement (TEAC/ABTS assay). A common cell-free antioxidant protocol involves: Dissolve ABTS in ultrapure water (7.4 mM, 0.4 mL) and mix with potassium persulfate (K₂S₂O₈) stock solution (2.6 mM, 1.43 mL). Allow the mixture to stand at room temperature in the dark for 12 hours to generate ABTS radical cations (ABTS⁺). Dilute the solution 50-fold with anhydrous ethanol. Add antioxidant samples and measure the decrease in absorbance at 734 nm to assess free radical scavenging capacity.
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| Toxicity/Toxicokinetics |
Based on available safety data, ABTS diammonium salt is classified as an irritant. Hazard classifications include Eye Irritant (Category B). The compound is unstable in solution and should be prepared fresh. General handling precautions include avoiding contact with eyes, skin, and inhalation of dust. Reagent-grade ABTS diammonium salt is strictly intended for research and diagnostic use only (e.g., ELISA), not for human therapeutic applications. Storage recommendations: 4°C, sealed, away from moisture; in solvent at -80°C for up to 6 months or at -20°C for up to 1 month.
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| References |
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| Additional Infomation |
ABTS diammonium salt is a water-soluble chromogenic substrate widely used for detecting peroxidase activity due to its stable oxidation products [1][2]. The oxidation reaction of peroxidase with ABTS diammonium salt depends on H₂O₂, and the resulting colored product can be used to quantitatively analyze enzyme activity or antigen-antibody binding events [1][2]. In the diagnosis of schistosomiasis, ELISA methods based on ABTS diammonium salt have advantages over traditional substrates, including faster reaction speed and lower background noise [1]. Recombinant DyP from Pleurotus erythrorhizon exhibits higher catalytic efficiency for ABTS diammonium salt than other common peroxidase substrates [2].
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| Molecular Formula |
C18H16N4O6S4.2(NH4)
|
|---|---|
| Molecular Weight |
548.68
|
| Exact Mass |
548.064
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| Elemental Analysis |
C, 39.40; H, 4.41; N, 15.32; O, 17.50; S, 23.37
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| CAS # |
30931-67-0
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| Related CAS # |
28752-68-3 (Parent)
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| PubChem CID |
9570474
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| Appearance |
Light green to green solid powder
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| Boiling Point |
881.2ºC at 760mmHg
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| Flash Point |
486.7ºC
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| LogP |
5.478
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
34
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| Complexity |
883
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCN1/C(=N/N=C/2\\SC3=C(N2CC)C=CC(=C3)S(=O)(=O)O)/SC4=C1C=CC(=C4)S(=O)(=O)O.N.N
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| InChi Key |
ZTOJFFHGPLIVKC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H18N4O6S4/c1-3-21-13-7-5-11(31(23,24)25)9-15(13)29-17(21)19-20-18-22(4-2)14-8-6-12(32(26,27)28)10-16(14)30-18/h5-10H,3-4H2,1-2H3,(H,23,24,25)(H,26,27,28)
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| Chemical Name |
3-ethyl-2-[(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid
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| Synonyms |
ABTS diammonium salt; MFCD00010404; 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt; ABTS Diammonium;
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O : ≥ 50 mg/mL (~91.13 mM)
DMSO : ~20.83 mg/mL (~37.96 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.79 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 20.8 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. Solubility in Formulation 2: ≥ 2.08 mg/mL (3.79 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 20.8 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (3.79 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 50 mg/mL (91.13 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8226 mL | 9.1128 mL | 18.2256 mL | |
| 5 mM | 0.3645 mL | 1.8226 mL | 3.6451 mL | |
| 10 mM | 0.1823 mL | 0.9113 mL | 1.8226 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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