概述

  • 产品名称Anti-Niemann Pick C1抗体
    参阅全部 Niemann Pick C1 一抗
  • 描述
    兔多克隆抗体to Niemann Pick C1
  • 经测试应用适用于: IP, ICC/IF, WBmore details
  • 种属反应性
    与反应: Human
    预测可用于: Rabbit, Chimpanzee, Macaque Monkey, Gorilla, Orangutan
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1250 to the C-terminus of Human Niemann Pick C1.

  • 阳性对照
    • This antibody gave a positive signal in Human Adrenal Gland tissue lysate as well as the following whole cell lysates: HepG2; WI38; HeLa; A431. It also gave a positive signal in HepG2 cell line.

性能

应用

Our Abpromise guarantee covers the use of ab108921 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
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 200 kDa (predicted molecular weight: 142 kDa).

靶标

  • 功能Involved in the intracellular trafficking of cholesterol. May play a role in vesicular trafficking in glia, a process that may be crucial for maintaining the structural and functional integrity of nerve terminals.
  • 疾病相关Defects in NPC1 are the cause of Niemann-Pick disease type C1 (NPDC1) [MIM:257220]. A lysosomal storage disorder that affects the viscera and the central nervous system. It is due to defective intracellular processing and transport of low-density lipoprotein derived cholesterol. It causes accumulation of cholesterol in lysosomes, with delayed induction of cholesterol homeostatic reactions. Niemann-Pick disease type C1 has a highly variable clinical phenotype. Clinical features include variable hepatosplenomegaly and severe progressive neurological dysfunction such as ataxia, dystonia and dementia. The age of onset can vary from infancy to late adulthood. An allelic variant of Niemann-Pick disease type C1 is found in people with Nova Scotia ancestry. Patients with the Nova Scotian clinical variant are less severely affected.
  • 序列相似性Belongs to the patched family.
    Contains 1 SSD (sterol-sensing) domain.
  • 结构域A cysteine-rich N-terminal domain and a C-terminal domain containing a di-leucine motif necessary for lysosomal targeting are critical for mobilization of cholesterol from lysosomes.
  • 翻译后修饰Glycosylated.
  • 细胞定位Late endosome membrane. Lysosome membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Niemann Pick C1 protein precursor antibody
    • Niemann Pick disease, type C1 antibody
    • Niemann-Pick C1 protein antibody
    • NPC antibody
    • NPC1 antibody
    • NPC1_HUMAN antibody
    see all

Anti-Niemann Pick C1 antibody 图像

  • Niemann Pick C1 was immunoprecipitated using 0.5mg HeLa whole cell extract, 5µg of Rabbit polyclonal to Niemann Pick C1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab108921.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 142kDa; Niemann Pick C1, non specific bands - 55kDa: We are unsure as to the identity of this extra band.

  • All lanes : Anti-Niemann Pick C1 antibody (ab108921) at 1 µg/ml (Milk Blocking - 3%)

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : Human adrenal normal tissue lysate - total protein (ab29249)
    Lane 3 : WI-38 whole cell lysate (ab3960)
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 142 kDa
    Observed band size : 200 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 42 kDa,60 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes
    Niemann Pick C1 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • ICC/IF image of ab108921 stained HepG2 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 (ab108921, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) HeLa cells at 5µg/ml, and in 100% methanol fixed (5 min) HepG2 cells at 5µg/ml.

Anti-Niemann Pick C1 antibody (ab108921)参考文献

This product has been referenced in:
  • Desai TM  et al. IFITM3 restricts influenza A virus entry by blocking the formation of fusion pores following virus-endosome hemifusion. PLoS Pathog 10:e1004048 (2014). WB ; Human . Read more (PubMed: 24699674) »

See 1 Publication for this product

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Unfortunately, I don't think that any of the http://www.abcam.com/index.html?t=158346&pt=1 we have in our catalog is suitable for the detection of the yeast Niemann-Pick type C-related protein 1 (http://www.unip...

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