Anti-activated Notch1抗体(ab8925)

概述

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

    Synthetic peptide corresponding to activated Notch1 aa 1755-1767 (intracellular). The epitope is only exposed after gamma secretase cleavage and is not accessible in the uncleaved form.
    Sequence:

    VLLSRKRRRQHGQC


    (Peptide available as ab730)

  • 阳性对照
    • Transiently transfected 293 cells expressing recombinant myc-tagged mouse Notch constructs (see image)
  • 常规说明

    Notch is synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase (S1 cleavage) 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 (S2 cleavage) 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 (S3 cleavage) to release the intracellular domain (NICD) from the membrane.

性能

应用

Our Abpromise guarantee covers the use of ab8925 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
IHC-P 1/200.
IHC-Fr Use at an assay dependent concentration. PubMed: 18371421
ICC/IF Use at an assay dependent concentration.
WB 1/500. Detects a band of approximately 80 kDa.Can be blocked with Human activated Notch1 peptide (ab730). 80kDa band is a cleavage product of Notch 1. There is also a contaminating band present at 100 kDa. The identity of this band is unknown.
Flow Cyt Use at an assay dependent concentration. PubMed: 19171875

ab171870 - 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. Represses neuronal and myogenic differentiation. May enhance HIF1A function by sequestering HIF1AN away from HIF1A.
  • 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 aortic valve disease 1 (AOVD1) [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). Following endocytosis, 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. Monoubiquitination at Lys-1759 is required for activation by gamma-secretase cleavage, it promotes interaction with AAK1, which stabilizes it. Deubiquitination by EIF3F is necessary for nuclear import of activated Notch.
    Hydroxylated at Asn-1955 by HIF1AN. Hydroxylated at Asn-2022 by HIF1AN (By similarity). Hydroxylation reduces affinity for HI1AN and may thus indirectly modulate negative regulation of NICD.
  • 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
    • hN1 antibody
    • Neurogenic locus Notch homolog protein 1 antibody
    • NICD antibody
    • NOTC1_HUMAN antibody
    • Notch 1 antibody
    • Notch 1 intracellular domain antibody
    • Notch homolog 1 translocation associated (Drosophila) antibody
    • notch1 antibody
    • TAN1 antibody
    • Translocation-associated notch protein TAN-1 antibody
    see all

Anti-activated Notch1 antibody 图像

  • ab8925 staining actived Notch1 in the NIH3T3 fibroblast cell line from Mouse embryo by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 0.1% and blocked with 10% serum for 30 minutes at 24°C. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C. An Alexa Fluor®488-conjugated Donkey anti-rabbit IgG polyclonal(1/500) was used as the secondary antibody.

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  • This image is courtesy of Stacey Huppert, Ma. Xenia U. Garcia (lab of Raphael Kopan)

    Rabbit polyclonal (ab8925) at 1/500, against myc-tagged transiently transfected mouse Notch constructs in 293 cells.

    Lane M: Mol wt markers
    Lane 1 No transfection
    Lane 2 N1 (mouse deleted extracellular domain)-myc
    Lane 3 N1 (mouse intracellular domain)-myc
    Lane 4 N2 (mouse deleted extracellular domain)-myc
    Lane 5 N2 (mouse intracellular domain)-myc

    Lane 6 N3 (mouse deleted extracellular domain)-myc
    Lane 7 N3 (mouse intracellular domain)-myc
    Lane 8 N4 (mouse deleted extracellular domain)-myc
    Lane 9 N4 (mouse intracellular domain)-myc
    Lane 10 N1 (mouse deleted extracellular domain)(V to G)-myc

    Rabbit polyclonal (ab8925) at 1/500, against myc-tagged transiently transfected mouse Notch constructs in 293 cells.

    Lane M: Mol wt markers

    Lane 1 No transfection

    Lane 2 N1 (mouse deleted extracellular domain)-myc

    Lane 3 N1 (mouse intracellular domain)-myc

    Lane 4 N2 (mouse deleted extracellular domain)-myc

  • ab8925 (1:500) diluted staining activated Notch1 in human ovarian carcinoma using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of activated Notch 1 in nuclear/nucleolar compartments of the ovarian cortex.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • ab8925 staining activated Notch 1 in murine dermis (including follicle) by Immunohistochemistry (Frozen sections). Tissue was fixed with paraformaldehyde and permeabilized using 0.1% Triton. Samples were then blocked with 10% serum for 1 hour at 19°C followed by incubation with the primary antibody at a 1/100 dilution for 16 hours at 4°C. An Alexa Fluor® conjugated donkey anti-rabbit polyclonal was used as secondary antibody at a 1/200 dilution. Counterstain DAPI (blue).

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Anti-activated Notch1 antibody (ab8925)参考文献

This product has been referenced in:
  • Nishio M  et al. Dysregulated YAP1/TAZ and TGF-ß signaling mediate hepatocarcinogenesis in Mob1a/1b-deficient mice. Proc Natl Acad Sci U S A 113:E71-80 (2016). WB ; Mouse . Read more (PubMed: 26699479) »
  • Dou GR  et al. Myeloid-Specific Blockade of Notch Signaling Attenuates Choroidal Neovascularization through Compromised Macrophage Infiltration and Polarization in Mice. Sci Rep 6:28617 (2016). IHC-FoFr ; Mouse . Read more (PubMed: 27339903) »

See all 42 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (mcf7)
Specification mcf7
Fixative acetone/methanol 50/50
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
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Abcam user community

Verified customer

提交于 Apr 06 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - nuclear (MCF-7)
Loading amount 25 µg
Specification MCF-7
Treatment Mock or EDTA 5mM for 10mins
Gel Running Conditions Reduced Denaturing (8-12% gradient)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Verified customer

提交于 Mar 27 2013

Thank you for your enquiry.

I can confirm that ab8925 antibody is sold as whole antiserum. Unpurified antibodies, such as those sold as whole antiserum will not have a concentration stated on the datasheet. Antibody concentration is usually de...

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Regarding the activated Notch1 antibody ab8925, this antibody is validated and guaranteed to work in IHC (frozen and paraffin sections), ICC, WB, and Flow cytometry u...

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Application Western blot
Sample Mouse Tissue lysate - other (brain)
Gel Running Conditions Non-reduced Denaturing (10% acrylamide)
Loading amount 25 µg
Treatment 25 uM DAPT
Specification brain
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Ms. Victoria Lladó

Verified customer

提交于 Oct 16 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (hepatocytes)
Permeabilization Yes - 0.1% Triton X-100 in TBS for 5-10 minutes
Specification hepatocytes
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 23°C
Fixative Paraformaldehyde
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提交于 Sep 11 2015

Application IHC - Wholemount
Sample Zebrafish Embryo (48hpf)
Specification 48hpf
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提交于 Mar 23 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Dakocytomation X0909 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH6.0
Sample Mouse Tissue sections (human melanoma cell line injected s.c. in mouse.)
Specification human melanoma cell line injected s.c. in mouse.
Permeabilization Yes - 0.25%Triton-X100
Fixative Formaldehyde
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Hongwei Shao

Verified customer

提交于 May 02 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C
Sample Mouse Cell (NIH3T3)
Specification NIH3T3
Permeabilization Yes - 0.1% Triton
Fixative Paraformaldehyde
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Verified customer

提交于 Jan 02 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (NIH3T3)
Specification NIH3T3
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
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Abcam user community

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提交于 Apr 05 2013

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