Recombinant Mouse ASAHL Protein, CF

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R&D Systems Recombinant Proteins and Enzymes
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Product Details

Recombinant Mouse ASAHL Protein, CF Summary

Product Specifications

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Measured by its ability to hydrolyze the substrate palmitoylethanolamide into palmitate and ethanolamine. The specific activity is >100 pmol/min/µg, as measured under the described conditions.
Mouse myeloma cell line, NS0-derived mouse ASAHL/N-acylethanolamine-hydrolyzing Acid Amidase protein
Val33-Ser362, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Predicted Molecular Mass
38 kDa
50 kDa, 33 kDa and 19 kDa, reducing conditions

Product Datasheets

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


Formulation Supplied as a 0.2 μm filtered solution in MES and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

  • Assay Buffer: 0.1 M Sodium Acetate, 0.1% (v/v) NP-40 substitute (Fluka, Catalog # 74385), pH 4.0
  • Recombinant Mouse ASAHL/N‑acylethanolamine-hydrolyzing Acid A (rmASAHL) (Catalog # 4886-AH)
  • Palmitoyl Ethanolamide (PEA) (Tocris, Catalog # 0879), 25 mM stock in dimethyl formamide
  • Dithiothreitol (DTT) (Sigma, Catalog # D0632), 1 M stock in deionized water
  • Sodium Hydroxide
  • beta -mercaptoethanol (Sigma, Catalog # M7154)
  • 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. Dilute PEA to 50 µM in Assay Buffer. Dissolve 10 µL of 25 mM stock into 4.99 mL of Assay Buffer (Note: Preheat assay buffer to
    37 °C and vortex for 30 seconds to completely solubilize the PEA).
  2. Dilute rmASAHL to 1.25 µg/mL in Assay Buffer.
  3. Set up reactions in 1.5 mL microtubes. Mix 200 µL of 50 µM PEA, 50 µL of 1.25 µg/mL rmASAHL, and 2.5 µL of 1 M DTT.
  4. Incubate reaction tubes at 37 °C for 1 hour.
  5. Prepare an o-PA solution. Mix 3.84 mL of 0.2 M Sodium Hydroxide, 4 µL beta -mercaptoethanol, and 160 µL 50 mg/mL o-PA.
  6. Add 250 µL of the o-PA mixture (step 5) to all reaction vials. Mix well and incubate at room temperature for 10 minutes.
  7. Create a control vial by combining, in this order, 250 µL o-PA mixture (step 5), 50 µL of 1.25 µg/mL rmASAHL, and 200 µL of 50 µM PEA.
  8. Load in a plate 200 µL in duplicate of reaction mixtures and control.
  9. Read at excitation and emission wavelengths of 330 nm and 450 nm (top read), respectively, in endpoint mode.
  10. Calculate specific activity (Average duplicates):

     Specific Activity (pmol/min/µg) =

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

     *Adjusted for control
     **Derived using calibration standard ethanolamine (Sigma, Catalog # E9508).

Per Well:
  • rmASAHL: 0.025 µg
  • PEA: 20 µM
  • o-PA: 1 mg/mL
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.


Background: ASAHL/N-acylethanolamine-hydrolyzing Acid Amidase

The mouse ASAHL gene encodes N-acylethanolamine-hydrolyzing Acid Amidase (NAAA), a fatty acid amidase with maximal activity at acidic pH (1). NAAA hydrolyzes a number of N-acyl ethanolamines, including N-myristoyl-, N-stearoyl-, N-oleoyl-, and N-arachidonoyl, but is most active against N-palmitoylethanolamine (2). NAAA is a member of the choloylglycine hydrolase family of enzymes, and is structurally similar to acid ceramidase (1). NAAA is both a lysosomal and a secreted enzyme, and like acid ceramidase, has been observed to be proteolytically processed during maturation (1). Through its amidase activity, ASAHL may play a role in the termination of the actions of a variety of N-acylethanolamides (3). NAAA can be distinguished from anandamide amidohydrolase by its lack of inhibition by methyl arachidonoyl fluorophosphonate (2).

  1. Tsuboi, K. et al. (2005) J. Biol. Chem. 280:11082.
  2. Ueda, N. et al. (2001) J. Biol. Chem. 276:35552.
  3. Sun, Y. X. et al. (2005) Biochim. Biophys. Acta 1736:211.
Entrez Gene IDs
27163 (Human); 67111 (Mouse); 497009 (Rat)
Alternate Names
Acid ceramidase-like protein; ASAHL; ASAH-like protein; EC 3.5.1.-; NAAA; N-acylethanolamine acid amidase; Nacylethanolaminehydrolyzing Acid Amidase; N-acylethanolamine-hydrolyzing acid amidase; N-acylsphingosine amidohydrolase (acid ceramidase)-like; N-acylsphingosine amidohydrolase-like; PLT


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