Buffer Formulation	PBS, pH7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300.
Traits	Freeze-dried powder
Purity	> 95%
Isoelectric Point	7.2
Applications	Cell culture; Activity Assays.

ACTIVITY TEST

Sphingomyelin phosphodiesterase 1 (SMPD1), also known as acid sphingomyelinase (ASM), belongs to the sphingomyelin phosphodiesterase family. The protein Converts sphingomyelin to ceramide. ASM also has phospholipase C activities toward 1,2-diacylglycerolphosphocholine and 1,2-diacylglycerolphosphoglycerol.
Thus, the recombinant human ASM activity was measured by its ability to hydrolyze 2-N-Hexadecanoylamino-4-nitrophenylphosphorylcholine (HDA-PC) to p-Nitrophenol. The reaction was performed in 50 mM MES, 0.5 µM ZnCl2, pH 7.5 ( Assay Buffer), initiated by addition 50 μL of various concentrations of ASM (diluted with Assay Buffer) to 50 µL of 1 mM Substrate HDA-PC ( 50 mM stock solution in methanol, diluted with Assay Buffer). Incubated at room temperature for 20 minutes in the dark, then read at a wavelength of 405 nm.

One unit of enzyme activity is defined as the 1μg of enzyme required to convert 1pmol of HDA-PC to p-Nitrophenol in 1min at 37°C. The specific activity of recombinant human ASM is >1000 pmol/min/µg.

 

USAGE

 

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

 

STORAGE

 

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

 

STABILITY

 

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

 

Image


Figure. SDS-PAGE

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