Anti-Acid sphingomyelinase antibody [mAbcam74281] (ab74281)

Overview

  • Product nameAnti-Acid sphingomyelinase antibody [mAbcam74281]
    See all Acid sphingomyelinase primary antibodies
  • Description
    Mouse monoclonal [mAbcam74281] to Acid sphingomyelinase
  • Tested applicationsWB, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Acid sphingomyelinase.

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HepG2; A431; MCF7; HeLa; THP1.
  • General notesWe can conjugate this antibody to FITC for you (please see ab150251 for details).

Properties

Applications

Our Abpromise guarantee covers the use of ab74281 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 70 kDa).
Flow Cyt Use 1µg for 106 cells.

Target

  • FunctionConverts sphingomyelin to ceramide. Also has phospholipase C activities toward 1,2-diacylglycerolphosphocholine and 1,2-diacylglycerolphosphoglycerol. Isoform 2 and isoform 3 have lost catalytic activity.
  • Involvement in diseaseDefects in SMPD1 are the cause of Niemann-Pick disease type A (NPDA) [MIM:257200]; also known as Niemann-Pick disease classical infantile form. It is an early-onset lysosomal storage disorder caused by failure to hydrolyze sphingomyelin to ceramide. It results in the accumulation of sphingomyelin and other metabolically related lipids in reticuloendothelial and other cell types throughout the body, leading to cell death. Niemann-Pick disease type A is a primarily neurodegenerative disorder characterized by onset within the first year of life, mental retardation, digestive disorders, failure to thrive, major hepatosplenomegaly, and severe neurologic symptoms. The severe neurological disorders and pulmonary infections lead to an early death, often around the age of four. Clinical features are variable. A phenotypic continuum exists between type A (basic neurovisceral) and type B (purely visceral) forms of Niemann-Pick disease, and the intermediate types encompass a cluster of variants combining clinical features of both types A and B.
    Defects in SMPD1 are the cause of Niemann-Pick disease type B (NPDB) [MIM:607616]; also known as Niemann-Pick disease visceral form. It is a late-onset lysosomal storage disorder caused by failure to hydrolyze sphingomyelin to ceramide. It results in the accumulation of sphingomyelin and other metabolically related lipids in reticuloendothelial and other cell types throughout the body, leading to cell death. Clinical signs involve only visceral organs. The most constant sign is hepatosplenomegaly which can be associated with pulmonary symptoms. Patients remain free of neurologic manifestations. However, a phenotypic continuum exists between type A (basic neurovisceral) and type B (purely visceral) forms of Niemann-Pick disease, and the intermediate types encompass a cluster of variants combining clinical features of both types A and B. In Niemann-Pick disease type B, onset of the first symptoms occurs in early childhood and patients can survive into adulthood.
  • Sequence similaritiesBelongs to the acid sphingomyelinase family.
    Contains 1 saposin B-type domain.
  • Cellular localizationLysosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • Acid sphingomyelinase antibody
    • ASM antibody
    • ASM_HUMAN antibody
    • aSMase antibody
    • NPD antibody
    • Smpd1 antibody
    • Sphingomyelin phosphodiesterase 1 acid lysosomal antibody
    • Sphingomyelin phosphodiesterase antibody
    see all

Anti-Acid sphingomyelinase antibody [mAbcam74281] images

  • All lanes : Anti-Acid sphingomyelinase antibody [mAbcam74281] (ab74281) at 5 µg/ml

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 70 kDa
    Observed band size : 65 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute
  • Overlay histogram showing HeLa cells stained with ab74281 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab74281, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-Acid sphingomyelinase antibody [mAbcam74281] (ab74281)

This product has been referenced in:
  • Simonis A  et al. Differential activation of acid sphingomyelinase and ceramide release determines invasiveness of Neisseria meningitidis into brain endothelial cells. PLoS Pathog 10:e1004160 (2014). Human . Read more (PubMed: 24945304) »

See 1 Publication for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (gel concentration 4-12%)
Sample Rat Tissue lysate - whole (blood vessel)
Specification blood vessel
Blocking step Milk as blocking agent for 6 hour(s) and 0 minute(s) · Concentration: 5%
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Verified customer

Submitted May 01 2014

Thank you for your enquiry.

Unfortunately, we have no further cross reactivity data, the immuogen blast (using UniProt) doesn't give any data for these species and we have only ever tested this in human lysates in WB and human cell lines in I...

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Thank you for contacting us and pointing out that this information was missing. I have contacted the laboratory and am happy to confirm that this antibody is of isotype IgG2a and light chain kappa. The last production of this antibody was subsequen...

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