Recombinant Mouse ASAH2 Protein, CF


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Product Details

Reactivity MuSpecies Glossary
Applications Enzyme Activity

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Recombinant Mouse ASAH2 Protein, CF Summary

Details of Functionality
Measured by its ability to hydrolyze the substrate C12:0 ceramide into sphingosine and dodecanoic acid. The specific activity is > 3,000 pmol/min/µg, as measured under the described conditions.
Mouse myeloma cell line, NS0-derived mouse ASAH2/N-acylsphingosine Amidohydrolase-2 protein
Thr34-Thr756, with an N-terminal 6-His tag
Accession # NP_061300
Accession #
N-terminal Sequence
Protein/Peptide Type
Recombinant Enzymes
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.


Theoretical MW
81 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
113 kDa, reducing conditions
Read Publications using
3558-AH in the following applications:

Packaging, Storage & Formulations

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Supplied as a 0.2 μm filtered solution in MES, NaCl and Sodium Cholate.
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 25 mM MES, 150 mM NaCl, 1% (w/v) Sodium Cholate, pH 6.5
  • o-PA Buffer: 0.2 M NaOH, 0.1% beta -mercaptoethanol (v/v)
  • Recombinant Mouse ASAH2/N‑acylsphingosine Amidohydrolase-2 (rmASAH2) (Catalog # 3558-AH)
  • Substrate: C12-ceramide (Avanti Polar Lipids, Catalog # 860512P), 50 mM stock in Chloroform
  • o-Phthaldialdehyde (o-PA) (Sigma, Catalog # P0657), 50 mg/mL stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dissolve 10 µL of 50 mM stock of Substrate in 1.99 mL Assay Buffer for a 250 µM concentration. (Note: Preheat assay buffer to
    37 °C and vortex for 30 seconds to dissolve Substrate).
  2. Dilute rmASAH2 to 0.05 µg/mL in Assay Buffer.
  3. Combine 200 µL of 250 µM Substrate and 50 µL of 0.05 µg/mL rmASAH2. Include one blank containing 50 µL rmASAH2 and 200 µL Assay Buffer and another blank containing 200 µL Substrate and 50 µL Assay Buffer.
  4. Incubate at 37 °C for 1 hour.
  5. Stop reactions by heating them at 95-100 °C for 5 minutes.
  6. Dilute o-PA to 2 mg/mL in o-PA Buffer.
  7. Add 250 µL of the o-PA mixture to all reaction vials, including controls. Mix well.
  8. Incubate at room temperature for 10 minutes. Note: It is important not to deviate from this incubation time.
  9. Load 200 µL (in duplicate) of reaction mixtures and controls in a plate.
  10. Read at excitation and emission wavelengths of 330 nm and 450 nm (top read), respectively, in endpoint mode.
  11. Calculate specific activity using the following equation:

     Specific Activity (pmol/min/µg) =

Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU)
Incubation time (min) x amount of enzyme (µg)

     *Average duplicates, use the control with the higher RFU value to adjust fluorescence.
     **Derived using calibration standard Sphingosine (Avanti Polar Lipids, Catalog # 860490P).

Per Well:
  • rmASAH2: 0.001 µg
  • C12-ceramide: 100 μM
  • o-PA: 1 mg/mL


This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse ASAH2 Protein, CF

  • ASAH2 N-acylsphingosine amidohydrolase (non-lysosomal ceramidase) 2
  • ASAH2
  • BCDase
  • HNAC1
  • LCDase
  • Nacylsphingosine Amidohydrolase2
  • NCDase
  • N-CDase


The mouse ASAH2 gene encodes acylsphingosine amidohydrolase-2, also known as neutral ceramidase. Neutral ceramidase is a type II integral membrane protein that can be cleaved to produce a soluble secreted protein (1). The enzyme is abundant in the brush border membranes of the intestine, but is also expressed in tissues such as kidney, brain and liver (2, 3). A major physiological function of neutral ceramidase is the metabolism of dietary sphingolipids, but the enzyme may also be involved in the generation of messenger molecules such as sphingosine and sphingosine 1-phosphate (3).

  1. Tani, M. et al. (2003) J. Biol. Chem. 278:10523.
  2. Kono, M. et al. (2006) J. Biol. Chem. 281:7324.
  3. Mitsutake, S. et al. (2001) J. Biol. Chem. 276:26249.

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Publications for ASAH2/N-acylsphingosine Amidohydrolase-2 (3558-AH)(2)

We have publications tested in 1 confirmed species: Mouse.

We have publications tested in 2 applications: Bioassay, In Vivo.

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Gene Symbol Asah2