| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
RNA (binds to residual RNA in reticulocytes)
DNA (binds to DNA, also used as a probe) [1] DNA and RNA targets (when conjugated to oligonucleotide probes) [2] DNA (when used as a gel staining agent) [3] |
|---|---|
| ln Vitro |
Upon binding to RNA, thiazole orange (compound 6) exhibits a several thousand-fold fluorescence enhancement. Its extinction coefficient (ε) with RNA is 3.8 × 10⁴ M⁻¹cm⁻¹, and the quantum yield (Φ) with RNA is 0.2, giving a “brightness” (Φ•ε) of approximately 8000. The fluorescence enhancement upon adding RNA is 3000-fold. [1]
Thiazole orange shows saturation of nucleic acid binding sites typical of strongly binding intercalating dyes. Saturation of RNA was achieved with a ratio of one dye to ten base pairs, and saturation of DNA was achieved with a ratio of one dye to two base pairs. [1] When conjugated to oligonucleotide probes (2'-OMe-(ORN3) AE-TO_Q6), thiazole orange showed up to a 44-fold fluorescence enhancement (F_ds/F_ss) against DNA targets. For example, 2'-OMe-(ORN3) AE-TO_Q6 gave an F_ds/F_ss of 33.9 (integrated 510-650 nm) or 43.9 (510-555 nm). The fluorescence quantum yield of AE-TO_Q6 in 2'-OMe-(ORN3) against a DNA target was 0.318, and in 2'-OMe-(ORN1) was 0.580. Single-stranded 2'-OMe RNA TO-oligonucleotides had low background fluorescence (Φ = 0.013-0.022). [2] Thiazole orange enables detection of DNA in agarose gels with a detection limit of approximately 1-2 ng/lane, comparable to ethidium bromide and a common blue-light-detectable commercial dye. [3] |
| Enzyme Assay |
Fluorescence titration of RNA and DNA with thiazole orange was performed. The concentration of RNA was 1.86 × 10⁻⁵ M bases, and the concentration of DNA was 3.25 × 10⁻⁶ M base pairs. Excitation and emission wavelengths were 510 nm and 530 nm, respectively. Fluorescence units were arbitrary. Saturation of RNA was achieved at a dye-to-base pair ratio of 1:10, and saturation of DNA at a ratio of 1:2. [1]
UV melting experiments were performed to determine duplex stability (Tm) of thiazole orange-labeled oligonucleotides (ODNs and 2'-OMe ORNs) with complementary DNA or RNA targets. Samples were measured in a buffer containing 10 mM phosphate and 200 mM NaCl at pH 7.0. DNA concentrations for Tm studies were 3.0 μM of probe and 3.3 μM of target strand. Tm values are an average of 4 |
| Cell Assay |
Flow cytometric analysis of whole blood stained with thiazole orange was performed. Whole blood (5 μL) was added to 1 mL of diluted dye solution (3 × 10⁻⁷ M) in phosphate-buffered saline containing 0.002 M EDTA and 0.02% sodium azide, and incubated at room temperature for 1 hour. An unstained control (blood in buffer without dye) was used. A minimum of 10,000 cells was collected at a flow rate between 500 to 1,000 cells per second. Fluorescence was measured with a 530/30-nm bandpass interference filter after excitation with 200 mW of 488 nm radiation from an argon-ion laser. Reticulocytes were identified as events to the right of a marker set using the unstained control histogram. Leukocytes were excluded by gating on forward scatter vs. fluorescence. [1]
Fluorescence emission spectroscopy of thiazole orange-modified ODNs (0.25 μM probe, 0.28 μM target) was performed in buffer containing 10 mM phosphate and 200 mM NaCl at pH 7.0, at 20°C. Excitation was at 484 nm, and emission was measured at 510 nm (slit width 7 nm). F_ds/F_ss ratios were calculated based on integrated fluorescence intensity (510-650 nm). [2] For cell membrane permeability testing, a suspension of human peripheral mononuclear cells (10 μL, 2×10⁷ cells/mL) was mixed with a solution of the dye in phosphate-buffered saline (10 μL, 10⁻⁵ M). A wet mount was examined under a light microscope and for fluorescence. Thiazole orange was determined to be membrane permeable because all cells were fluorescent. [1] |
| Toxicity/Toxicokinetics |
Thiazole orange is less mutagenic than ethidium bromide (3-4x less mutagenic in the Ames test) and can be generally disposed of with common chemical waste. It is considered to have a favorable safety profile. [3]
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| References | |
| Additional Infomation |
Thiazole orange is a cyanine dye composed of a thiazole orange cation [1-methyl-4-[(3-methyl-1,3-benzothiazole-2(3H)-ylidene)methyl]quinolinium] and a p-toluenesulfonate anion. It is a fluorescent dye. It contains the thiazole orange cation.
Thiazole orange is positively charged; during agarose gel electrophoresis it will migrate in the opposite direction of DNA. For samples run far down the gel, the dye may separate from smaller DNA fragments, resulting in weak staining; in such instances, post-electrophoresis staining is recommended. [3] Thiazole orange can be excited with UV light (for use as a direct replacement for ethidium bromide) or with blue light (e.g., 470 nm or 488 nm) to prevent DNA damage. Blue light does not cause skin damage or DNA damage, though intense blue light may still be harmful to eyes; an amber emission filter (~560 nm longpass) should be used for visualization. [3] Thiazole orange was originally named and evaluated as a reticulocyte dye (compound 6 in Table 1). It showed a correlation coefficient of 0.97 compared to manual determination of reticulocyte percentage with new methylene blue across several blood samples. [1] |
| Molecular Formula |
C19H17N2S+.C7H7O3S-
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|---|---|
| Molecular Weight |
476.61036
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| Exact Mass |
476.122
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| CAS # |
107091-89-4
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| PubChem CID |
6438345
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| Appearance |
Pink to red solid powder
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| Melting Point |
270ºC (dec.)(lit.)
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| LogP |
5.307
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
33
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| Complexity |
628
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=CC=C(C=C1)S(=O)(=O)[O-].CN\1C2=CC=CC=C2S/C1=C\C3=CC=[N+](C4=CC=CC=C34)C
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| InChi Key |
ACOJCCLIDPZYJC-UHFFFAOYSA-M
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| InChi Code |
InChI=1S/C19H17N2S.C7H8O3S/c1-20-12-11-14(15-7-3-4-8-16(15)20)13-19-21(2)17-9-5-6-10-18(17)22-19;1-6-2-4-7(5-3-6)11(8,9)10/h3-13H,1-2H3;2-5H,1H3,(H,8,9,10)/q+1;/p-1
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| Chemical Name |
4-methylbenzenesulfonate;(2Z)-3-methyl-2-[(1-methylquinolin-1-ium-4-yl)methylidene]-1,3-benzothiazole
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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) |
MEOH : 25 mg/mL (~52.45 mM)
DMSO : ~25 mg/mL (~52.45 mM) H2O : < 0.1 mg/mL |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (2.62 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 12.5 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: ≥ 1.25 mg/mL (2.62 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 12.5 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0982 mL | 10.4908 mL | 20.9815 mL | |
| 5 mM | 0.4196 mL | 2.0982 mL | 4.1963 mL | |
| 10 mM | 0.2098 mL | 1.0491 mL | 2.0982 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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