Anti-Acid sphingomyelinase抗体(ab83354)

概述

  • 产品名称Anti-Acid sphingomyelinase抗体
    参阅全部 Acid sphingomyelinase 一抗
  • 描述
    兔多克隆抗体to Acid sphingomyelinase
  • 经测试应用适用于: ICC/IF, WB, IHC-Pmore details
  • 种属反应性
    与反应: Human
    预测可用于: Mouse, Rat, Sheep, Rabbit, Horse, Guinea pig, Cow, Cat, Dog, Pig
  • 免疫原

    Synthetic peptide corresponding to a region within internal sequence amino acids 396-445 (INSTDPAGQL QWLVGELQAA EDRGDKVHII GHIPPGHCLK SWSWNYYRIV) of Human Acid sphingomyelinase (NP_000534).

  • 阳性对照
    • Fetal heart lysate.

性能

应用

Our Abpromise guarantee covers the use of ab83354 in the following tested applications.

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

应用 Ab评论 说明
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 70 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

靶标

  • 功能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.
  • 疾病相关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.
  • 序列相似性Belongs to the acid sphingomyelinase family.
    Contains 1 saposin B-type domain.
  • 细胞定位Lysosome.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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 图像

  • Anti-Acid sphingomyelinase antibody (ab83354) at 1 µg/ml (in 5% skim milk / PBS buffer) + Human fetal heart lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size : 70 kDa
    Observed band size : 65 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 58 kDa. We are unsure as to the identity of these extra bands.
  • ICC/IF image of ab83354 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab83354, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • IHC image of ab83354 staining in human normal cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab83354, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Anti-Acid sphingomyelinase antibody (ab83354)参考文献

This product has been referenced in:
  • Brauweiler AM  et al. Th2 Cytokines Increase Staphylococcus aureus Alpha Toxin-Induced Keratinocyte Death through the Signal Transducer and Activator of Transcription 6 (STAT6). J Invest Dermatol N/A:N/A (2014). Read more (PubMed: 24468745) »
  • Defour A  et al. Dysferlin regulates cell membrane repair by facilitating injury-triggered acid sphingomyelinase secretion. Cell Death Dis 5:e1306 (2014). WB ; Mouse . Read more (PubMed: 24967968) »

See all 4 Publications for this product

Product Wall

Abreviews
Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Mouse Cell lysate - whole cell (Primary hepatocytes)
Specification Primary hepatocytes
Treatment recombinant adenovirus carrying Asmase gene
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 May 29 2013

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