| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
Octamer-binding transcription factor 3/4 (Oct3/4) [1]
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| ln Vitro |
O4I2 not only stimulates Oct3/4 expression by activating its promoter site, but also stabilizes the Oct3/4 protein in HEK293 cells[1]. O4I2 substantially activates Oct3/4 even at 5 µM after 72 hrs treatment on the translational level[1]. O4I2 (20 µM) accumulates Oct3/4 in NCCIT happened already 2 hours after incubation, but the measurable Sox2 increase emerges much later, implying compound-mediated activation of Oct3/4 might be different between embryonic kidney HEK293 cells and embryonal cancer NCCIT cells [1].
In HEK293 cells transfected with Oct3/4 promoter-driven luciferase reporter plasmid, O4I2 (1-10 μM) dose-dependently activated Oct3/4 promoter activity, with a maximal 4.2-fold increase at 5 μM compared to vehicle control. This indicated potent induction of Oct3/4 transcriptional activity[1] - In P19 embryonic carcinoma cells and mouse embryonic stem cells (mESCs), O4I2 (0.5-20 μM) upregulated Oct3/4 mRNA expression by 3.5-5.0-fold (RT-PCR) and protein levels by 2.8-3.6-fold (Western blot) at 10 μM. It also moderately increased the expression of other pluripotency-associated markers (Sox2, Nanog) by 1.5-2.0-fold[1] - In differentiated mESCs (induced by retinoic acid), O4I2 (10 μM) reversed the downregulation of Oct3/4, restoring its mRNA and protein levels to ~70% of undifferentiated mESC levels, suggesting potential pluripotency-retaining activity[1] |
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| ln Vivo |
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| Cell Assay |
Immunofluorescence[1]
Cell Types: HF (human fibroblasts) cells. Tested Concentrations: 5-20 µM. Incubation Duration: 5 days. Experimental Results: Successfully detected co-expression of Oct3/4 and Sox2, as well as Nanog and Klf4. Oct3/4 promoter luciferase reporter assay: HEK293 cells were seeded in 96-well plates and co-transfected with Oct3/4 promoter-luciferase plasmid and Renilla luciferase plasmid (internal control). After 24 hours, O4I2 (0.1 μM, 1 μM, 5 μM, 10 μM, 20 μM) was added, and cells were cultured for another 48 hours. Luciferase activity was measured using a dual-luciferase assay system, with relative luciferase activity (firefly/Renilla) reflecting Oct3/4 promoter activation[1] - Pluripotency marker expression assay: P19 cells and mESCs were seeded in 6-well plates and treated with O4I2 (0.5 μM, 5 μM, 10 μM, 20 μM) for 72 hours. For RT-PCR, total RNA was extracted, reverse-transcribed to cDNA, and amplified with specific primers for Oct3/4, Sox2, and Nanog. For Western blot, cell lysates were prepared, subjected to electrophoresis, transferred to membranes, and probed with antibodies against Oct3/4, Sox2, Nanog, and β-actin (loading control)[1] - Differentiated mESC rescue assay: mESCs were induced to differentiate with retinoic acid for 4 days. O4I2 (10 μM) was added during the last 3 days of differentiation. Oct3/4 mRNA and protein levels were detected by RT-PCR and Western blot, respectively, and compared to undifferentiated and vehicle-treated differentiated mESCs[1] |
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| Animal Protocol |
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| References | |||
| Additional Infomation |
O4I2 is a derivative of ethyl 2-((4-chlorophenyl)amino)thiazole-4-carboxylate and has been shown to be a potent inducer of the pluripotency transcription factor Oct3/4[1]. Its core mechanism is to activate the Oct3/4 promoter and upregulate the expression of Oct3/4 mRNA and protein, which may help maintain or restore the pluripotency of stem cells[1]. O4I2 has shown dose-dependent activity in various cell models (HEK293, P19, mESCs) and has shown cross-regulatory effects on other pluripotency markers (Sox2, Nanog), which supports its potential as a tool compound for stem cell biology research[1]. The thiazole skeleton of O4I2 is crucial to its Oct3/4 induction activity, providing a structural basis for the development of novel pluripotency regulators[1].
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| Molecular Formula |
C12H11CLN2O2S
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|---|---|---|
| Molecular Weight |
282.75
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| Exact Mass |
282.022
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| CAS # |
165682-93-9
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| Related CAS # |
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| PubChem CID |
53428670
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
408.1±51.0 °C at 760 mmHg
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| Flash Point |
200.6±30.4 °C
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| Vapour Pressure |
0.0±1.0 mmHg at 25°C
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| Index of Refraction |
1.637
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| LogP |
3.83
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
18
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| Complexity |
285
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
ULUBAPWNHROTEU-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C12H11ClN2O2S/c1-2-17-11(16)10-7-18-12(15-10)14-9-5-3-8(13)4-6-9/h3-7H,2H2,1H3,(H,14,15)
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| Chemical Name |
ethyl 2-((4-chlorophenyl)amino)thiazole-4-carboxylate
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| Synonyms |
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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 |
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| 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) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (8.84 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5367 mL | 17.6835 mL | 35.3669 mL | |
| 5 mM | 0.7073 mL | 3.5367 mL | 7.0734 mL | |
| 10 mM | 0.3537 mL | 1.7683 mL | 3.5367 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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