ERNT
ERNT is a fluorescent probe for monitoring endoplasmic reticulum (ER) polarity and detecting endoplasmic reticulum stress (ERS) during the progression of dynamic liver injury. ERNT functions based on a donor-π-acceptor (D-π-A) structure with an intramolecular charge transfer (ICT) effect. ERNT has multiple excitation/emission pairs: for in vitro cell imaging, the green channel has Ex/Em = 488/500-550 nm, and the red channel has Ex/Em = 488/570-620 nm; for in vivo liver imaging, Ex/Em = 520/620 nm; for solvent optical tests, Ex = 470 nm, with the maximum emission peak at 548 nm in low-polarity toluene and 668 nm in high-polarity DMSO. ERNT can evaluate the efficacy of hepatoprotective interventions by detecting changes in ER polarity, and also exhibits excellent photostability and low cytotoxicity.
For research use only. We do not sell to patients.
- CAS No.: 2767443-23-0
- Formula: C39H31N3O4S
- Molecular Weight:637.75
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
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Biological Activity
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Chemical Information
- Protocols
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
Description
In Vitro
Guideline
(The following is a recommended experimental protocol for guidance only, and adjustments are required based on your specific needs)[1].
1. Preparation of Working Solution
1.1 Concentration recommendations: 10 μM; 15 μM.
2. Staining Procedures
2.1 Sample types:
2.1.1 Adherent cells (HepG2).
2.1.2 Liver tissue samples: Isolate the liver organ and rinse it with PBS (pH 7.4).
2.2 Incubation conditions:
2.2.1 Adherent cells: Incubate with 10 μM ERNT (with continuous irradiation for 0, 5, 10, 15 and 20 min).
2.2.2 Liver tissue samples: Incubate with 15 μM ERNT.
2.3 Washing procedures:
2.3.1 Liver tissue samples: Rinse three times with PBS (pH 7.4) before staining.
3. Control Group Setup
3.1 Set up untreated adherent cells as the control group.
3.2 Set up untreated liver-injured mice as the control group.
3.3 Set up normal liver tissue as the reference group.
4. Detection and Analysis
4.1 Instruments: Fluorescence imaging system; In vivo imaging system.
4.1.1 Adherent cells: Green channel: Ex = 488 nm, Em = 500-550 nm; Red channel: Ex = 488 nm, Em = 570-620 nm.
4.1.2 Liver tissue samples: Ex = 520 nm, Em = 620 nm; Ex = 570 nm, Em = 710 nm.
4.2 Result analysis:
4.2.1 Changes in fluorescence intensity:
4.2.1.1 Higher fluorescence intensity indicates lower endoplasmic reticulum (ER) polarity; lower fluorescence intensity indicates higher ER polarity.
4.2.1.2 Fluorescence intensity decreases gradually from normal liver tissue to fatty liver, liver fibrosis, liver cirrhosis and finally liver cancer.
4.2.1.3 Compared with the untreated control group, the fluorescence intensity of the drug-treated liver injury group increases.
4.2.1.4 The fluorescence intensity ratio of green channel to red channel (Igreen/Ired) in normal cells is higher than that in ER stress-induced cells.
4.2.2 Fluorescence localization: Mainly localized to the endoplasmic reticulum.
4.2.3 Color changes: Emits green and red fluorescence in cells; emits yellow to red fluorescence in liver tissue.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
Chemical Information
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CAS No. 2767443-23-0
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Molecular Weight 637.75
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Formula C39H31N3O4S
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SMILES
CC1=CC=C(C=C1)S(=O)(NCCN2C(C3=CC=C(C4=CC=CC(C2=O)=C34)C5=CC=C(C=C5)N(C6=CC=CC=C6)C7=CC=CC=C7)=O)=O
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Protocols
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Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
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Mammalian live/dead viability and cytotoxicity staining
Live/dead viability and cytotoxicity staining assays are based on the simultaneous detection of intracellular esterase activity in metabolically active (viable) cells and membrane integrity loss in non-viable cells. In commonly used dual-staining approaches, membrane-permeant fluorogenic substrates are converted by intracellular esterases into fluorescent products in live cells, while impermeant DNA-binding dyes selectively enter cells with compromised plasma membranes and label nucleic acids in dead or dying cells, enabling discrimination between viable and non-viable populations by fluorescence microscopy or flow cytometry.
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Cell Viability Determination by MTT Colorimetric Assay
The following protocol uses the MTT colorimetric assay as a classic literature-established method for assessing cell viability/metabolic activity in cultured mammalian cells. MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] is reduced by metabolically active cells to a colored formazan product; the amount of formazan is quantified spectrophotometrically and provides an indirect measure of metabolically active viable cells. Importantly, MTT reduction reflects cellular oxidoreductase/metabolic activity rather than an absolute direct count of living cells, so changes in cellular metabolism can alter the signal independently of cell number.
Purity & Documentation
References
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
ERNT Related Classifications
HY-D3104 Related Classifications
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