概述

  • 产品名称Anti-Notch1抗体
    参阅全部 Notch1 一抗
  • 描述
    兔多克隆抗体to Notch1
  • 经测试应用适用于: ICC/IF, IHC-Fr, IP, IHC-P, WB, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 2500 to the C-terminus of Human Notch1.

    (Peptide available as ab68580.)

  • 阳性对照
    • This antibody gave a positive signal in the following whole cell lysates: HeLa, HepG2, HEK293, SK N SH and Jurkat. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2.

性能

应用

Our Abpromise guarantee covers the use of ab65297 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 5 µg/ml.
IHC-Fr 1/100.
IP Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 272 kDa).
Flow Cyt Use at an assay dependent concentration. PubMed: 24136221ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

靶标

  • 功能Functions as a receptor for membrane-bound ligands Jagged1, Jagged2 and Delta1 to regulate cell-fate determination. Upon ligand activation through the released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus. Affects the implementation of differentiation, proliferation and apoptotic programs. May be important for normal lymphocyte function. In altered form, may contribute to transformation or progression in some T-cell neoplasms. Involved in the maturation of both CD4+ and CD8+ cells in the thymus. May be important for follicular differentiation and possibly cell fate selection within the follicle. During cerebellar development, may function as a receptor for neuronal DNER and may be involved in the differentiation of Bergmann glia.
  • 组织特异性In fetal tissues most abundant in spleen, brain stem and lung. Also present in most adult tissues where it is found mainly in lymphoid tissues.
  • 疾病相关Defects in NOTCH1 are a cause of bicuspid aortic valve (BAV) [MIM:109730]. A common defect in the aortic valve in which two rather than three leaflets are present. It is often associated with aortic valve calcification and insufficiency. In extreme cases, the blood flow may be so restricted that the left ventricle fails to grow, resulting in hypoplastic left heart syndrome.
  • 序列相似性Belongs to the NOTCH family.
    Contains 5 ANK repeats.
    Contains 36 EGF-like domains.
    Contains 3 LNR (Lin/Notch) repeats.
  • 翻译后修饰Synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase in the trans-Golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin dependent gamma-secretase to release a notch-derived peptide containing the intracellular domain (NICD) from the membrane.
    Phosphorylated.
    O-glycosylated on the EGF-like domains. Contains both O-linked fucose and O-linked glucose.
    Ubiquitinated; undergoes 'Lys-29'-linked polyubiquitination catalyzed by ITCH.
  • 细胞定位Cell membrane and Nucleus. Following proteolytical processing NICD is translocated to the nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 9930111A19Rik antibody
    • AOS5 antibody
    • AOVD1 antibody
    • hN1 antibody
    • Lin-12 antibody
    • LIN12 antibody
    • Mis6 antibody
    • Motch A antibody
    • mT14 antibody
    • Neurogenic locus notch homolog protein 1 antibody
    • Neurogenic locus notch protein homolog antibody
    • NICD antibody
    • NOTC1_HUMAN antibody
    • Notch 1 antibody
    • Notch 1 intracellular domain antibody
    • NOTCH antibody
    • Notch gene homolog 1 (Drosophila) antibody
    • Notch homolog 1, translocation-associated (Drosophila) antibody
    • NOTCH, Drosophila, homolog of, 1 antibody
    • notch1 antibody
    • TAN1 antibody
    • Translocation associated notch homolog antibody
    • Translocation associated notch protein TAN 1 antibody
    • Translocation-associated notch protein TAN-1 antibody
    • xotch antibody
    see all

Anti-Notch1 antibody 图像

  • Notch1 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Rabbit polyclonal to Notch1 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, Hek293 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 ab65297.

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

    Band: 120kDa; Notch1

  • All lanes : Anti-Notch1 antibody (ab65297) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : SK N SH (Human neuroblastoma) Whole Cell Lysate
    Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 272 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 160 kDa,50 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
    The band observed at 120 kDa could potentially be a truncated form of Notch1, following cleavage by furin-like protease. This band is also observed with other commercially available antibodies to Notch1.
  • IHC image of Notch1 staining in Human Hippocampus FFPE section, performed on a 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 ab65297, 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
  • ICC/IF image of ab65297 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 ab65297 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.

  • ab65297 staining Notch1 (green) in Mouse embryo tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in a 0.5% citrate buffer. Samples were incubated with primary antibody (1/50 in 5% Goat serum) for 2 hours. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody. Red - Myh6/7, Blue - nuclei (DAPI).

    See Abreview

  • ab65297 staining Notch1 (green) in Mouse embryo tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 1% Triton X-100 and blocked with 5% serum for 2 hours at room temperature. Samples were incubated with primary antibody (1/100 in 5% Goat serum) for 2 hours. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody. Red - Myh6/7, Blue - nuclei (DAPI).

    See Abreview

Anti-Notch1 antibody (ab65297)参考文献

This product has been referenced in:
  • Hu GH  et al. Delta-like ligand 4 (Dll4) predicts the prognosis of clear cell renal cell carcinoma, and anti-Dll4 suppresses tumor growth in vivo. Int J Clin Exp Pathol 7:2143-52 (2014). WB ; Human . Read more (PubMed: 24966922) »
  • Liu H  et al. Single-cell clones of liver cancer stem cells have the potential of differentiating into different types of tumor cells. Cell Death Dis 4:e857 (2013). Flow Cyt ; Human . Read more (PubMed: 24136221) »

See all 2 Publications for this product

Product Wall

Thank you for your reply.

I can confirm that ab65297 and ab52627 both have immunogens that are also located within the NICD (amino acid 1754 to 2555).

Therefore they also bind the region you are interested in. They areas specific as...

Read More

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