製品の概要

  • 製品名
    Anti-Niemann Pick C1 antibody
    Niemann Pick C1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Niemann Pick C1
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: IP, ICC/IF, WBmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: 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.

  • ポジティブ・コントロール
    • 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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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).

ターゲット情報

  • 機能
    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.
  • 関連疾患
    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.
  • 配列類似性
    Belongs to the patched family.
    Contains 1 SSD (sterol-sensing) domain.
  • ドメイン
    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.
  • 翻訳後修飾
    Glycosylated.
  • 細胞内局在
    Late endosome membrane. Lysosome membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

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

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : NPC1 (Niemann Pick C1) knockout HAP1 whole cell lysate
    Lane 3 : HEK293 whole cell lysate
    Lane 4 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 142 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab108921 observed at 200 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab108921 was shown to recognize Niemann Pick C1 in wild-type HAP1 cells as signal was lost at the expected MW in NPC1 (Niemann Pick C1) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NPC1 (Niemann Pick C1) knockout samples were subjected to SDS-PAGE. Ab108921 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • 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
    All lanes : 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.

参考文献

This product has been referenced in:
  • Mishra S  et al. Accumulation of cholesterol and increased demand for zinc in serum-deprived RPE cells. Mol Vis 22:1387-1404 (2016). WB . Read more (PubMed: 28003730) »
  • 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 all 2 Publications for this product

レビューと Q&A

Thank you for your inquiry.

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...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

登録