cis-Ned19
Based on 1 Customer Validation
cis-Ned19 is an irreversible nicotinic acid adenine dinucleotide phosphate (NAADP) antagonist. cis-Ned19 localizes to lysosomes and endolysosomal vesicles, stains two-pore calcium channels and fluorescently labels NAADP receptors (Ex/Em = 365/410 nm).
For research use only. We do not sell to patients.
- Purity: 99.25%
- CAS No.: 1137264-00-6
- Formula: C30H31FN4O3
- Molecular Weight:514.59
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Storage:
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Purity & Documentation
- References
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Help & FAQs
All Calcium Channel Isoforms
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Biological Activity
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Calcium Channel |
Guidelines (The following is our recommended protocol; this protocol serves as a guideline only and should be modified to suit your specific needs.)
1. Preparation of Stock and Working Solutions
a. Prepare a 10 mM cis-Ned19 stock solution in anhydrous DMSO.
b. Dilute the stock solution using pre-warmed, serum-free cell culture medium or PBS to prepare a 10 μM cis-Ned19 working solution.
Note: Please adjust the concentration of the cis-Ned19 working solution according to your specific experimental requirements, and prepare it immediately before use.
2. Cell Staining (Suspension Cells)
a. Centrifuge to harvest the cells, then wash twice with PBS (5 minutes per wash). Adjust the cell density to 1×106 cells/mL.
b. Add 1 mL of the cis-Ned19 working solution and incubate at room temperature for 10 minutes.
c. Centrifuge at 400 × g for 3–4 minutes, and discard the supernatant.
d. Wash the cells twice with PBS (5 minutes per wash).
e. Resuspend the cells in 1 mL of serum-free medium or PBS, then observe using a fluorescence microscope or flow cytometer.
3. Cell Staining (Adherent Cells)
a. Culture adherent cells on sterile coverslips.
b. Remove the coverslips from the culture medium and aspirate any excess medium.
c. Add the working solution, gently agitating to ensure it completely covers the cells, and incubate at room temperature for 10 minutes.
d. Aspirate the dye working solution, then wash 2–3 times with culture medium (5 minutes per wash). Observe using a fluorescence microscope or flow cytometer.
Note: If analysis via flow cytometry is required, the cells must be detached using trypsin digestion and resuspended prior to staining.
cis-NED19 (10 μM) binds to sites localized in endolysosomes of human umbilical vein endothelial cells[1].
cis-Ned19 (10 min) colocalizes significantly with the lysosomal marker LysoTracker but not with endoplasmic reticulum or mitochondrial markers in cultured human umbilical vein endothelial cells[2].
cis-Ned19 inhibits Ca2+ release (IC50 = 800 nM) and [32P]NAADP binding (IC50 = 15 μM)[3].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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CAS No. 1137264-00-6
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Appearance Solid
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Molecular Weight 514.59
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Formula C30H31FN4O3
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Color White to off-white
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SMILES
O=C([C@@H]1CC2=C([C@H](C3=CC=C(OC)C(CN4CCN(C5=CC=CC=C5F)CC4)=C3)N1)NC6=C2C=CC=C6)O
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
Solvent & Solubility
DMSO : ≥ 100 mg/mL (194.33 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
* "≥" means soluble, but saturation unknown.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Purity & Documentation
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Data Sheet (280 KB)
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SDS (393 KB)
- English - EN (393 KB)
- Français - FR (393 KB)
- Deutsch - DE (393 KB)
- Norwegian - NO (393 KB)
- Español - ES (393 KB)
- Swedish - SV (393 KB)
- Italian - IT (393 KB)
- Korean - KR (393 KB)
- Portuguese - PT (393 KB)
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Handling Instructions (2659 KB)
References
[1]. Zharkich IL, et al. Suppression of Histamine-Induced Relaxation of Rat Aorta and Calcium Signaling in Endothelial Cells by Two-Pore Channel Blocker trans-NED19 and Hydrogen Peroxide. Izv Akad Nauk Ser Biol. 2016;(4):430-438. [Content Brief]
[2]. Avdonin PV, et al. Involvement of two-pore channels in hydrogen peroxide-induced increase in the level of calcium ions in the cytoplasm of human umbilical vein endothelial cells. Dokl Biochem Biophys. 2017;474(1):209-212. [Content Brief]
[3]. Naylor E, et al. Identification of a chemical probe for NAADP by virtual screening. Nat Chem Biol. 2009 Apr;5(4):220-6. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 1.9433 mL | 9.7165 mL | 19.4329 mL | 48.5824 mL |
| 5 mM | 0.3887 mL | 1.9433 mL | 3.8866 mL | 9.7165 mL | |
| 10 mM | 0.1943 mL | 0.9716 mL | 1.9433 mL | 4.8582 mL | |
| 15 mM | 0.1296 mL | 0.6478 mL | 1.2955 mL | 3.2388 mL | |
| 20 mM | 0.0972 mL | 0.4858 mL | 0.9716 mL | 2.4291 mL | |
| 25 mM | 0.0777 mL | 0.3887 mL | 0.7773 mL | 1.9433 mL | |
| 30 mM | 0.0648 mL | 0.3239 mL | 0.6478 mL | 1.6194 mL | |
| 40 mM | 0.0486 mL | 0.2429 mL | 0.4858 mL | 1.2146 mL | |
| 50 mM | 0.0389 mL | 0.1943 mL | 0.3887 mL | 0.9716 mL | |
| 60 mM | 0.0324 mL | 0.1619 mL | 0.3239 mL | 0.8097 mL | |
| 80 mM | 0.0243 mL | 0.1215 mL | 0.2429 mL | 0.6073 mL | |
| 100 mM | 0.0194 mL | 0.0972 mL | 0.1943 mL | 0.4858 mL |
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