C12 NBD Sphingomyelin

• CAT Number: I043069
• CAS Number: 254117-01-6
• Molecular Formula: C41H73N6O9P
• Molecular Weight: 825.03
• Purity: ≥95%
• Synonyms: [(E,2S,3R)-3-hydroxy-2-[12-[(4-nitro-2,1,3-benzoxadiazol-7-yl)amino]dodecanoylamino]octadec-4-enyl] 2-(trimethylazaniumyl)ethyl phosphate
• InChI: InChI=1S/C41H73N6O9P/c1-5-6-7-8-9-10-11-12-13-15-18-21-24-27-38(48)36(34-55-57(52,53)54-33-32-47(2,3)4)43-39(49)28-25-22-19-16-14-17-20-23-26-31-42-35-29-30-37(46(50)51)41-40(35)44-56-45-41/h24,27,29-30,36,38,48H,5-23,25-26,28,31-34H2,1-4H3,(H2-,42,43,45,49,52,53)/b27-24+/t36-,38+/m0/s1
• InChIKey: FCQMKLRMKNYONQ-YPGNEAGUSA-N
• SMILES: CCCCCCCCCCCCCC=CC(C(COP(=O)([O-])OCC[N+](C)(C)C)NC(=O)CCCCCCCCCCCNC1=CC=C(C2=NON=C12)[N+](=O)[O-])O

C12 NBD sphingomyelin is an active derivative of sphingomyelin (HY-113498) that is tagged with fluorescent C12 nitrobenzoxadiazole (C12 NBD). C12 NBD sphingomyelin can be used as a sphingomyelinase substrate for studying the metabolism and transport of sphingomyelins (Ex=470 nm, Em=525 nm)[1].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)[1].
Assay for Sphingolipid-Degrading Enzymes (EGCase Ⅱ, SCDase and SMase):
1. Incubate amounts of enzymes with 0.1 nM dye at 37 ℃ for indicated times under following conditions.
(1). 10 mM sodium acetate buffer (pH5.0) containing 0.2% Triton X-100 for EGCase.
(2) 25 mM sodium phosphate buffer (pH 6.0) containing 0.1% Triton X-100 for SCDase.
(3) 25 mM sodium phosphate buffer (pH 7.0) containing 0.2% Triton X-100 for SMase.
2. After incubation, the solvent is evaporated and the residue is dried, dissolved in 10 μL of chloroform/methanol (2:1) and analyzed by TLC using chloroform/methanol/0.02% CaCl2 (5:4:1, v/v) as the developing solvent.
3. Degradation products and remaining substrates are separated by TLC and quantified with a chromatoscanner (excitation 470 nm, emission 525 nm) for fluorescence-labeled substrates.

Reference

[1]. Nakagawa, et al. Preparation of fluorescence-labeled GM1 and sphingomyelin by the reverse hydrolysis reaction of sphingolipid ceramide N-deacylase as substrates for assay of sphingolipid-degrading enzymes and for detection of sphingolipid-binding proteins. J. Biochem. 126(3), 601-611 (1999).
 [Content Brief]