Anti-Acid sphingomyelinase antibody [mAbcam74281] (ab74281)
Key features and details
- Mouse monoclonal [mAbcam74281] to Acid sphingomyelinase
- Suitable for: WB, Flow Cyt (Intra)
- Reacts with: Human
- Isotype: IgG2a
Overview
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Product name
Anti-Acid sphingomyelinase antibody [mAbcam74281]
See all Acid sphingomyelinase primary antibodies -
Description
Mouse monoclonal [mAbcam74281] to Acid sphingomyelinase -
Host species
Mouse -
Tested applications
Suitable for: WB, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human Acid sphingomyelinase aa 1-100 conjugated to keyhole limpet haemocyanin.
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Positive control
- This antibody gave a positive signal in the following whole cell lysates: HepG2; A431; MCF7; HeLa; THP1.
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General notes
We can conjugate this antibody to FITC for you (please see ab150251 for details).
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
mAbcam74281 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgG2a -
Light chain type
kappa -
Research areas
Associated products
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Assay kits
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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WB | (1) |
Use a concentration of 5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 70 kDa).
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Flow Cyt (Intra) |
Use 1µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
Use a concentration of 5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 70 kDa). |
Flow Cyt (Intra)
Use 1µg for 106 cells. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
Target
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Function
Converts 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 disease
Defects 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 similarities
Belongs to the acid sphingomyelinase family.
Contains 1 saposin B-type domain. -
Cellular localization
Lysosome. - Information by UniProt
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Database links
- Entrez Gene: 6609 Human
- Omim: 607608 Human
- SwissProt: P17405 Human
- Unigene: 498173 Human
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Alternative names
- Acid sphingomyelinase antibody
- ASM antibody
- ASM_HUMAN antibody
see all
Images
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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
All lanes : 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
The predicted molecular weight of Acid Sphingomyelinase protein is 70 kDa. However, the protein sequence contains a 46-residue signal sequence at the amino-terminal, which could explain the band observed. -
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.
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab74281 has been referenced in 5 publications.
- Lv Y et al. Synergism of rMV-Hu191 with cisplatin to treat gastric cancer by acid sphingomyelinase-mediated apoptosis requiring integrity of lipid raft microdomains. Gastric Cancer 24:1293-1306 (2021). PubMed: 34251544
- Su YT et al. Hyperbaric Oxygen Treatment Ameliorates Hearing Loss and Auditory Cortex Injury in Noise Exposed Mice by Repressing Local Ceramide Accumulation. Int J Mol Sci 20:N/A (2019). PubMed: 31547176
- Peters S et al. Neisseria meningitidis Type IV Pili Trigger Ca2+-Dependent Lysosomal Trafficking of the Acid Sphingomyelinase To Enhance Surface Ceramide Levels. Infect Immun 87:N/A (2019). PubMed: 31160362
- Stephan M et al. Role of caspases in CD95-induced biphasic activation of acid sphingomyelinase. Oncotarget 8:20067-20085 (2017). PubMed: 28223543
- 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 . PubMed: 24945304