• Product name
    Anti-activated Notch1 antibody
    See all activated Notch1 primary antibodies
  • Description
    Rabbit polyclonal to activated Notch1
  • Host species
  • Specificity
    ab52301 detects endogenous levels of fragment of activated Notch 1 resulting from cleavage adjacent to Val1744.
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, IHC-P, WB, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human activated Notch1 aa 1720-1770.


    Database link: P46531

  • Positive control
    • Extracts from NIH 3T3 cells, treated with etoposide (25µM, 1hour). IHC-P: Human brain tissue
  • General notes

    ab52301 is cleaved at Val 1744



Our Abpromise guarantee covers the use of ab52301 in the following tested applications.

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

Application Abreviews Notes
IHC-Fr Use at an assay dependent concentration. PubMed: 24098462
ICC/IF Use at an assay dependent concentration. PubMed: 20680961
IHC-P Use at an assay dependent concentration.
WB 1/500 - 1/1000. Detects a band of approximately 95 kDa (predicted molecular weight: 272 kDa).
ELISA 1/10000.


  • Function
    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.
  • Tissue specificity
    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.
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the NOTCH family.
    Contains 5 ANK repeats.
    Contains 36 EGF-like domains.
    Contains 3 LNR (Lin/Notch) repeats.
  • Post-translational
    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.
    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.
  • Cellular localization
    Cell membrane and Nucleus. Following proteolytical processing NICD is translocated to the nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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


  • Immunofluorescence analysis of frozen tissue sections (10µm) of intestine from sham operated rats (control group) or rats after Ischemic-Reperfusion (I/R) injury labeling Notch1 with ab52301. Tissue sections were frozen after being fixed with 4% formaldehyde, then incubated in 3% H2Ofor 30 minutes and then blocked with 5% BSA for 30 mins at room temperature. The sections were incubated overnight with ab52301 (PBS + 3% BSA). An appropriate  secondary antibody was then added. DAPI (in blue) was used to stain the nucleus.

  • All lanes : Anti-activated Notch1 antibody (ab52301) at 1/500 dilution

    Lane 1 : NIH 3T3 cell extract treated with etoposide
    (25µM, 1 hour)
    Lane 2 : NIH 3T3 cell extract treated with etoposide
    (25µM, 1 hour) with immunising peptide

    Predicted band size: 272 kDa
    Observed band size: 95 kDa
    why is the actual band size different from the predicted?

  • Immunohistochemical analysis of paraffin-embedded human brain tissue sections labeling activated Notch1 with ab52301 at a dilution of 1/100. The picture on the right is treated with the synthesized peptide. 


This product has been referenced in:
  • Liu W  et al. Mesenchymal stem cells alleviate the early brain injury of subarachnoid hemorrhage partly by suppression of Notch1-dependent neuroinflammation: involvement of Botch. J Neuroinflammation 16:8 (2019). Read more (PubMed: 30646897) »
  • Zhou H  et al. Paeonol reverses promoting effect of the HOTAIR/miR-124/Notch1 axis on renal interstitial fibrosis in a rat model. J Cell Physiol 234:14351-14363 (2019). Read more (PubMed: 30714138) »
See all 34 Publications for this product

Customer reviews and Q&As

1-5 of 5 Q&A


Ab52301 does detect the activated, intracellular domain of Notch 1 that forms after cleavage at Valine 1744 (murine) or Valine 1754 (human).

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Yes, the protease inhibitors may help, and to get bands in all lanes at 120 kDa, it may simply be necessary to develop the blot more: load more sample, use more antibody (for example 1/250), and incubate overnight at 4C in a solution of 1% BSA instead of 5%. This will probably not eliminate the band at 40 kDa (though using protease inhibitors when you collect your samples might), but you should still be able to obtain good data at 120 kDa, despitethe bandat 40 kDa. It likely represents an irrelevant protein that is much more abundant than Notch 1.

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Thank you for bringing this to our attention and for sending your protocol. I think your protocol is fine. I am a little concerned about the quality of the samples. From what you have described, there may be some degradation which would account for the smaller size bands and the absence of signal in some samples. Even though the actin band is more consistent, Notch may be more susceptible to proteolytic degradation than actin.

Can you please tell me how the skin samples are harvested and prepared for loading into the gel? You mentioned trying the antibody again with a new dilution, and that you saw no signal. Was this a new blot, or did you strip the original blot, and try re-staining it?

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Thank you for contacting us. We have a number of anti-Notch1 antibodies in our catalogue, however, none are specifically raised to detect the transmembrane domain. The transmembrane domain of human Notch1 is composed of residues 1736-1756 (SwissProt: P46531). We havean antibody which have been raised against an immunogen taken from residues 1755-1767, just outside the region of interest (https://www.abcam.com/activated-Notch1-antibody-ab8925.html). We also have an antibody which has been raised to specifically detect the cleavege site betweenglycine 1753 and valine 1754 (https://www.abcam.com/Notch1-antibody-Cleaved-Val1744-ab52301.html). This binds to the C-terminal cleavage product produced from this andit only recognises the cleaved form. Unfortunately these are the only antibodies I am able to suggest to your customer. I am sorry that I have not been of more help.

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Thank you for contacting Abcam. I have talked to the lab about whether ab52301 recognizes other bands. They have told me that the antibody can detect endogenous levels of fragment of activated Notch 1 resulting from cleavage adjacent to Val1754. The band at 120kD is the cleaved form, and the band at about 270kD is the full length form. Sometimes it can detect the full length form. The band that you are seeing at around 100kDa could be further breakdown products of your protein of interest or non-specific binding of the antibody. Also looking at some of our other Notch-1 antibodies (I am looking at ab65297), they also detect a (faint) band at ~100kDa in human kidney samples (in this case HEK cells). I have searched out catalogue for any lysates that you could use as a positive control but I have not found anything suitable. Please let me know if there is anything else I can help you with.

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