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  2. C12-NBD-ceramide

C12-NBD-ceramide 

Cat. No.: HY-141575
Handling Instructions

C12-NBD-ceramide is a fluorescent analogue of ceramide, it can be used as a substrate in ceramidase assays.

For research use only. We do not sell to patients.

C12-NBD-ceramide Chemical Structure

C12-NBD-ceramide Chemical Structure

CAS No. : 202850-01-9

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Description

C12-NBD-ceramide is a fluorescent analogue of ceramide, it can be used as a substrate in ceramidase assays[1][2].

In Vitro

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
In this experiment, C12-NBD-ceramide is used as a fluorescent nCDase substrate and reaction products will be separated by reverse-phase HPLC[2]:
1. 20 µM substrate, 1 nM nCDase in 75 mM NaCl, 25 mM HEPES (pH 8.0), and 0.4% Triton X-100, for 2 h at 37°C in a final volume of 100 µL.
2. Extract samples with 1:1 chloroform-methanol dries under nitrogen gas, and resuspends in 60 µL of HPLC mobile phase B. Separats reaction products by reverse-phase HPLC using a Spectra 3 µm C8SR column (3 μm particle, 3.0 × 150 mm).
3. Mobile phase A contains 0.2% formic acid and 1 mM ammonium formate in HPLC-grade water. Mobile phase B contains 0.2% formic acid (IL) and 1 mM ammonium formate in HPLC-grade methanol.
4. FRET-based assay using 20 µM substrate ES.173.cds, 50 ng of nCDase, 75 mM NaCl, 15 mM sodium phosphate (pH 7.4), and 0.3% Triton X-100 for 3 h in 96 black well plates.
5. Measure resorufin fluorescence (Ex=347 nm, Em=430 and 530 nm).

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

659.90

Formula

C36H61N5O6

CAS No.
SMILES

CCCCCCCCCCCCC/C=C/[[email protected]@H](O)[[email protected]](CO)NC(CCCCCCCCCCCNC1=CC=C([N+]([O-])=O)C2=NON=C12)=O

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Please store the product under the recommended conditions in the Certificate of Analysis.

Purity & Documentation
References
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Product Name:
C12-NBD-ceramide
Cat. No.:
HY-141575
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