NIR-ASM
NIR-ASM is a near-infrared fluorescent probe that can cross cell membranes and be activated by NQO1. NIR-ASM can distinguish NQO1-expressing cancer cells from normal cells via fluorescence microscopy and flow cytometry. NIR-ASM generates near-infrared fluorescence with a high signal-to-noise ratio in tumor models with NQO1 activity, enabling the detection of endogenous NQO1 activity in vivo. NIR-ASM is applicable to the research of lung cancer and breast cancer.
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- CAS No.: 2646628-35-3
- Formule: C33H35N3O3
- Masse moléculaire:521.65
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Stockage:
Please store the product under the recommended conditions in the Certificate of Analysis.
To place orders, for customer services and technical support, please contact: MedChemExpress USA
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Activité biologique
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Chemical Information
- Protocole
- Pureté et documentation
- Références
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Help & FAQs
Activité biologique
Description
IC50 & Target
[1]|
NQO1 |
In Vitro
NIR-ASM is an "on-type" fluorescent probe activatable by NQO1, suitable for cell-free biochemical assays. Its fluorescence response to NQO1 concentration shows a linear relationship within the range of 0.125 to 2.5 μg/mL, with a limit of detection of 0.191 μg/mL and a Km value of 33.82 μM for NQO1[1].
NIR-ASM selectively detects endogenous NQO1 activity in NQO1-positive A549 and NCI-H460 cancer cells, but fails to detect such activity in NQO1-negative IMR-90, HUVEC or untransfected MDA-MB-231 cells, and its activation strictly depends on the expression of NQO1[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
In Vivo
NIR-ASM (5 mg/kg; intravenous injection; single administration) selectively generates a near-infrared fluorescent signal in NQO1-positive orthotopic LLC lung tumors of C57BL/6 mice, with the signal peaking at 30 minutes post intravenous administration, while no signal is detected in the lungs of healthy mice[1].
NIR-ASM (5 mg/kg; 5 times per week; for 5 consecutive weeks) does not induce obvious toxicity or histological changes in healthy CD1 mice[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:athymic nude (nu/nu) (female, 4-6 weeks old, subcutaneous xenograft model)[1]
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Dosage:5 mg/kg
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Administration:i.v.; single dose
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Result:Reached a plateau in near-infrared fluorescence signal in the A549 tumor region at 30 minutes post-administration.
Detected no fluorescence signal in other major organs.
Confirmed fluorescence signal was restricted to the tumor via ex vivo imaging.
Detected NQO1 protein only in tumor lysates, not in other tissue lysates via Western blot analysis.
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Animal Model:C57BL/6 (female, orthotopic metastatic lung cancer model)[1]
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Dosage:5 mg/kg
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Administration:i.v.; single dose
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Result:Reached a maximum near-infrared fluorescence signal in the tumor-bearing lung region at 30 minutes post-administration.
Showed fluorescence signal intensity in tumor-bearing lungs more than 100-fold higher than in other major organs.
Confirmed strong fluorescence signal in tumor-bearing lungs with minimal signal in other organs via ex vivo imaging.
Detected no fluorescence signal in the lungs of non-tumor-bearing mice treated with NIR-ASM.
Chemical Information
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CAS No. 2646628-35-3
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Masse moléculaire 521.65
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Formule C33H35N3O3
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SMILES
CC1=C(C(C(C(C)(CC(NC2=CC=C(C=C2)/C=C/C3=C/C(CC(C)(C3)C)=C(C#N)/C#N)=O)C)=C(C1=O)C)=O)C
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Livraison
Room temperature in continental US; may vary elsewhere.
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Stockage
Please store the product under the recommended conditions in the Certificate of Analysis.
Protocole
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Flow Cytometry
Flow cytometry (FC) is a technique for high-speed, step-by-step quantitative analysis and sorting of single cells or other biological particles in a suspension by detecting labeled fluorescent signals.
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Flow cytometric DNA-content cell-cycle staining
Flow cytometric DNA-content cell-cycle staining measures the fluorescence intensity of DNA-bound fluorochromes in single cells or nuclei to estimate DNA content distributions, allowing assignment of populations to G0/G1, S, and G2/M phases by DNA histogram deconvolution. Propidium iodide (PI) intercalates into DNA, and PI fluorescence is proportional to cellular DNA content when staining is performed under conditions that make DNA accessible and minimize non-DNA signal. Cells with G2/M DNA content are expected to show approximately twice the fluorescence intensity of G0/G1 cells, while S-phase cells occupy intermediate fluorescence values. PI-based DNA-content analysis can also detect cells with fractional DNA content, often reported as sub-G1, when DNA fragmentation and extraction during staining reduce retained DNA signal in apoptotic cells. DAPI is an alternative DNA fluorochrome for univariate DNA-content analysis, while bivariate approaches combining DNA content with proliferation
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Protocol for Phospho-flow cytometry
Phospho-flow cytometry detects intracellular phosphorylated signaling proteins in single cells using phospho-specific antibodies after rapid fixation and permeabilization; the fluorescence intensity reflects phosphorylation state and therefore kinase-pathway activation, inhibition, or drug response in defined cell subsets. Unlike Western blot, phospho-flow preserves single-cell resolution and can measure signaling heterogeneity in cancer cells, primary immune cells, dissociated mouse tumors, macrophages, organoid-derived cells, and drug-screening samples when validated antibodies and fixation/permeabilization conditions are used.
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Breast Cancer Modeling
Breast cancer is a heterogeneous cancer, and it has been distinguished into four subtypes: luminal A, luminal B, HER2-positive and basal-like. Molecular mutations, epigenetic alterations, hormone exposure and immune microenvironment are related to the progression of breast cancer.
Pureté et documentation
Références
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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