Human Acid sphingomyelinase ELISA Kit (SMPD1) is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Acid sphingomyelinase (SMPD1) in Cell culture media, Heparin Plasma, Serum, EDTA Plasma samples.
Application | Reactivity | Dilution info | Notes |
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Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Converts sphingomyelin to ceramide (PubMed:12563314, PubMed:1840600, PubMed:18815062, PubMed:25339683, PubMed:25920558, PubMed:27659707, PubMed:33163980). Exists as two enzymatic forms that arise from alternative trafficking of a single protein precursor, one that is targeted to the endolysosomal compartment, whereas the other is released extracellularly (PubMed:20807762, PubMed:21098024, PubMed:9660788). However, in response to various forms of stress, lysosomal exocytosis may represent a major source of the secretory form (PubMed:12563314, PubMed:20530211, PubMed:20807762, PubMed:22573858, PubMed:9393854). In the lysosomes, converts sphingomyelin to ceramide (PubMed:20807762, PubMed:21098024). Plays an important role in the export of cholesterol from the intraendolysosomal membranes (PubMed:25339683). Also has phospholipase C activities toward 1,2-diacylglycerolphosphocholine and 1,2-diacylglycerolphosphoglycerol (PubMed:25339683). Modulates stress-induced apoptosis through the production of ceramide (PubMed:8706124). When secreted, modulates cell signaling with its ability to reorganize the plasma membrane by converting sphingomyelin to ceramide (PubMed:12563314, PubMed:17303575, PubMed:20807762). Secreted form is increased in response to stress and inflammatory mediators such as IL1B, IFNG or TNF as well as upon infection with bacteria and viruses (PubMed:12563314, PubMed:20807762, PubMed:9393854). Produces the release of ceramide in the outer leaflet of the plasma membrane playing a central role in host defense (PubMed:12563314, PubMed:20807762, PubMed:9393854). Ceramide reorganizes these rafts into larger signaling platforms that are required to internalize P. aeruginosa, induce apoptosis and regulate the cytokine response in infected cells (PubMed:12563314). In wounded cells, the lysosomal form is released extracellularly in the presence of Ca(2+) and promotes endocytosis and plasma membrane repair (PubMed:20530211). Sphingomyelin phosphodiesterase, processed form. This form is generated following cleavage by CASP7 in the extracellular milieu in response to bacterial infection (PubMed:21157428). It shows increased ability to convert sphingomyelin to ceramide and promotes plasma membrane repair (By similarity). Plasma membrane repair by ceramide counteracts the action of gasdermin-D (GSDMD) perforin (PRF1) pores that are formed in response to bacterial infection (By similarity). (Microbial infection) Secretion is activated by bacteria such as P. aeruginos, N. gonorrhoeae and others, this activation results in the release of ceramide in the outer leaflet of the plasma membrane which facilitates the infection. (Microbial infection) Secretion is activated by human coronaviruses SARS-CoV and SARS-CoV-2 as well as Zaire ebolavirus, this activation results in the release of ceramide in the outer leaflet of the plasma membrane which facilitates the infection. Isoform 2. Lacks residues that bind the cofactor Zn(2+) and has no enzyme activity. Isoform 3. Lacks residues that bind the cofactor Zn(2+) and has no enzyme activity.
ASM, SMPD1, Sphingomyelin phosphodiesterase, Acid sphingomyelinase, aSMase
Human Acid sphingomyelinase ELISA Kit (SMPD1) is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Acid sphingomyelinase (SMPD1) in Cell culture media, Heparin Plasma, Serum, EDTA Plasma samples.
Sample | n | mean | SD | C.V. |
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Sample Serum | n 8 | mean - | SD - | C.V. 4.9 |
Sample | n | mean | SD | C.V. |
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Sample Serum | n 0 | mean - | SD - | C.V. - |
Sample type | Average % | Range |
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Sample type Serum | Average % = 102 | Range |
Sample type EDTA Plasma | Average % = 103 | Range |
Sample type Cell culture media | Average % = 96 | Range |
Sample type Heparin Plasma | Average % = 98 | Range |
Human Acid sphingomyelinase ELISA kit (ab277075) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Acid sphingomyelinase protein in human serum, plasma - heparin, plasma - edta, and cell culture media. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Acid sphingomyelinase with 17.33 pg/mL sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
Acid Sphingomyelinase is an enzyme that converts sphingomyelin to ceramide. Mutations in Acid Sphingomyelinase are associated with Niemann-Pick disease A. A SNP in Acid Sphingomyelinase is a risk factor for Parkinson disease.
Acid sphingomyelinase (ASMase) also known as sphingomyelin phosphodiesterase 1 or NP is an enzyme involved in sphingolipid metabolism. ASMase has a mass of approximately 75 kDa and appears in lysosomes where it converts sphingomyelin to ceramide and phosphorylcholine. This enzyme is important in maintaining cellular lipid balance and signaling. Expression of ASMase occurs in various tissues such as the liver spleen and brain.
ASMase plays a role in membrane microdomain composition through its involvement in ceramide production. It participates in generating ceramide-enriched platforms that facilitate the clustering of signaling molecules. Ceramide acts as a second messenger in multiple cellular processes including apoptosis proliferation and inflammation. ASMase operates in the lysosomal lipid degradation pathway and connects with other lysosomal enzymes to modulate lipid turnovers such as glucosylceramidase affecting downstream cellular functions.
Sphingolipid metabolism involves ASMase. This enzyme participates in the ceramide signaling pathway influencing apoptosis and stress responses. Related proteins in this pathway include casein kinase II which phosphorylates ASMase and cathepsin D involved in the lysosomal degradation process. ASMase activity alters ceramide levels impacting pro-apoptotic and pro-survival signals mediated by related proteins in the cell signaling network.
ASMase deficiency connects to Niemann-Pick disease types A and B characterized by lipid accumulation in lysosomes. Mutations in the ASMase gene lead to impaired enzyme function resulting in excessive sphingomyelin storage and cell damage. The disorder links ASMase to proteins such as hexa-beta-N-acetylglucosaminidase which is affected in other lysosomal storage disorders. Research shows that ASMase activity also influences cardiovascular diseases by regulating ceramide and cholesterol levels in atherosclerotic lesions connecting it to inflammatory pathways involving adhesion molecules and cytokines.
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Example of human Acid Sphingomyelinase standard curve in Sample Diluent NS.
The Acid Sphingomyelinase standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Interpolated concentrations of native Acid Sphingomyelinase in human serum, plasma (heparin), and plasma (EDTA) samples.
The concentrations of Acid Sphingomyelinase were measured in duplicates, interpolated from the target standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma (heparin) 25%, and plasma (EDTA) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean target concentration was determined to be 5.04 ng/ml in serum, 4.63 ng/ml in plasma (heparin), and 3.75 ng/ml in plasma (EDTA).
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