WST-1

• CAT Number: R000824
• CAS Number: 150849-52-8
• Molecular Formula: C19H11IN5NaO8S2
• Molecular Weight: 651.34
• Purity: ≥95%
• Synonyms: sodium;4-[2-(4-iodophenyl)-3-(4-nitrophenyl)tetrazol-3-ium-5-yl]benzene-1,3-disulfonate
• InChI: InChI=1S/C19H12IN5O8S2.Na/c20-12-1-3-13(4-2-12)23-21-19(22-24(23)14-5-7-15(8-6-14)25(26)27)17-10-9-16(34(28,29)30)11-18(17)35(31,32)33;/h1-11H,(H-,28,29,30,31,32,33);/q;+1/p-1
• InChIKey: JUJBNYBVVQSIOU-UHFFFAOYSA-M
• SMILES: C1=CC(=CC=C1N2N=C(N=[N+]2C3=CC=C(C=C3)[N+](=O)[O-])C4=C(C=C(C=C4)S(=O)(=O)[O-])S(=O)(=O)[O-])I.[Na+]

WST-1 is a kind of water-soluble tetrazolium salt. WST induces the intracellular mitochondrial dehydrogenase to conduct NADH-dependent enzyme digestion reaction, releasing the water-soluble methyl benzene product. WST-1 can be used for the detection of cell proliferation and cytotoxicity, via the determination of the light absorption value at 450 nm[1].
Preparation of WST-1 working solution
1.1 Preparation of the stock solution
Dissolve 10 mg of WST-1 in 1 mL of DMSO to obtain 10 mg/mL of WST-1.
1.2 Preparation of WST-1 working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 5-10 μM of WST-1 working solution.
Note: Please adjust the concentration of WST-1 working solution according to the actual situation.

Cell staining
2.1 Cell preparation.For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of WST-1 working solution and 1-methoxy PMS (2 mM), and then incubate at room temperature for 2 h.
2.3 Use microplate reader detect 450/690 nm OD.

Reference

[1]. Munetaka ISHIYAMA, et al. A new sulfonated tetrazolium salt that produces a highly water-soluble formazan dye. Chem. Pharm. Bull. 1993, 41(6): 1118-1122.