Anti-Notch1 intracellular domain antibody (ab8387)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 intracellular domain antibody (ab8387)

    Key features and details

    • Rabbit polyclonal to Notch1 intracellular domain
    • Suitable for: IHC-P
    • Reacts with: Human
    • Isotype: IgG

    Overview

    • Product name

      Anti-Notch1 intracellular domain antibody

    • Description

      Rabbit polyclonal to Notch1 intracellular domain

    • Host species

      Rabbit

    • Specificity

      Notch 1 Does not recognise Notch 2

    • Tested Applications & Species

      Application Species
      IHC-P
      Human
      See all applications and species data

    • Immunogen

      Synthetic peptide:

      CQHSYSSPVDNTPSHQ

      , corresponding to amino acids 2488-2502 of Notch1 intracellular domain.
      Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

    • General notes

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

      One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

      Learn more here.

    Properties

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab8387 in the following tested applications.

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

    Guaranteed

    Tested applications are guaranteed to work and covered by our Abpromise guarantee.

    Predicted

    Predicted to work for this combination of applications and species but not guaranteed.

    Incompatible

    Does not work for this combination of applications and species.

    Application Species
    IHC-P
    Human
    Application Abreviews Notes
    IHC-P
    1/1000 - 1/5000.
    Notes
    IHC-P
    1/1000 - 1/5000.

    Target

    • 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.

    • 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 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.

    • Sequence similarities

      Belongs to the NOTCH family.
      Contains 5 ANK repeats.
      Contains 36 EGF-like domains.
      Contains 3 LNR (Lin/Notch) repeats.

    • Post-translational
      modifications

      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.

    • 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 antibody
        • notch1 antibody
        • TAN1 antibody
        • Translocation associated Notch protein TAN1 antibody
        • Translocation-associated notch protein TAN-1 antibody
        see all

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 intracellular domain antibody (ab8387)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 intracellular domain antibody (ab8387)

        ab8387 staining Notch1 in Human brain cerebellum tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde. Samples were incubated with primary antibody (1/500). Nuclear counter stain using hematoxylin (purple).

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (3)

      Publishing research using ab8387? Please let us know so that we can cite the reference in this datasheet.

      ab8387 has been referenced in 3 publications.

      • Castro Colabianchi AM  et al. Notch1 is asymmetrically distributed from the beginning of embryogenesis and controls the ventral center. Development 145:N/A (2018). PubMed: 29866901
      • Chu D  et al. Notch1 expression, which is related to p65 Status, is an independent predictor of prognosis in colorectal cancer. Clin Cancer Res 17:5686-94 (2011). PubMed: 21771876
      • Efstratiadis A  et al. Notch, Myc and breast cancer. Cell Cycle 6:418-29 (2007). PubMed: 17329972

      Customer reviews and Q&As

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      Question

      I bought anti-Notch1intra antibody (product n. ab8387) and tried it but it did not function well, too many bands and there was a band also at the right molecular weight in a mouse extract which should not be, because the antibody, as says the data sheet, should not cross-react with mouse. I used a classical protocol (5%milk in PBS with 0.2%Tween-20 which should be more stringent than BSA). So my question is: Do you have a picture of an experiment with Notch intra at the correct molecular weight made on human and mouse extracts in order that I can see what I should expect? Another question: how is it posible that this antibody recognizes only the Notch intra domain, since the longer forms of that protein should all contain the epitope at aa2788-2502?

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      Answer

      Thank you again for your email and patience while I have been trying to obtain additional information regarding this antibody. Unfortunately, there is no additional information regarding the testing of antibody, including a Western blot image. A colleague and I compared the human and mouse Notch1 intra sequences (amino acids 2488-2502) and there is 93% homology between the two sequences. Also, the epitope recognized by this antibody may be exposed in the cleaved form, but not in the full length form, even though the sequence is found in both. What dilution of ab8387 are you using? What is your secondary antibody and what dilution are you using? Have you done a secondary-only control to ensure that the secondary is not binding non-specifically? Also, what is your order number? Please rest assured that we guarantee our antibodies to work as stated on the online datasheets. I will do my best to troubleshoot this problem, and can offer you ca refund or replacement. Thanks you again, and I look forward to hearing from you.

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