SMPD1 (Sphingomyelin phosphodiesterase 1; also ASM) is a 70-72kD member of the sphingomyelinase family of enzymes. It is a monomeric lysosomal enzyme that converts sphingomyelin (a plasma membrane lipid) into ceramide through the removal of phosphorylcholine. This generates second messenger components that participate in signal transduction. Mature human SMPD1 is 583aa in length. It contains one saposin B-type domain (aa85-169) that likely interacts with sphingomyelin, and a catalytic region (aa 267-363). There are multiple isoforms. One is 57kD in size and results from proteolytic cleavage between Pro179-Pro197. Three others show aa substitutions; a 56aa substitution for (aa1-361), a two aa substitution for (aa363-629), a12aa substitution for (aa363-418). Over (aa62-628), human SMPD1 is 86% aa identical to mouse SMPD1.
Applications
Suitable for use in Direct ELISA, Immunoprecipitation and Neutralization. Other applications not tested.
Recommended Dilutions
Immunoprecipitation: 25ug/ml using conditioned cell culture medium spiked with Recombinant human SMPD1 Neutralization: Measured by its ability to neutralize Recombinant Human SMPD1 (0.5ug/ml) cleavage of the substrate 2-N-Hexadecanoylamino- 4-nitrophenylphosphorylcholine (HNPPC, 250uM). The Neutralization Dose (ND50) is typically 3ug/ml. Optimal dilutions to be determined by the researcher.
Hybridoma
Myeloma cells with B cells from Balb/c mice.
Storage and Stability
Lyophilized and reconstituted products are stable for 12 months after receipt at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Immunogen
Recombinant protein corresponding to His62-Pro628 from human SMPD1 isoform1, expressed in CHO cells.
Form
Supplied as a lyophilized powder from PBS, pH 7.4, 5% trehalose. Reconstitute with 200ul sterile PBS.
Purity
Purified by Protein A affinity chromatography from hybridoma culture supernatant.
Specificity
Recognizes human SMPD1. In direct ELISAs, no cross-reactivity with recombinant human SMPD3 and recombinant mouse SMPD1 is observed.