Recombinant
RabMAb

Recombinant Anti-Notch1 antibody [EP1238Y] (ab52627)

Rabbit recombinant monoclonal Notch1 antibody [EP1238Y]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Mouse, Cow, Human. Cited in 67 publication(s). Independently reviewed in 10 review(s).

Overview

  • Product name
    Anti-Notch1 antibody [EP1238Y]
    See all Notch1 primary antibodies
  • Description
    Rabbit monoclonal [EP1238Y] to Notch1
  • Host species
    Rabbit
  • Specificity
    80% identities with Notch 2 and 81% with Notch 3
  • Tested applications
    Suitable for: IHC-P, Flow Cyt, WB, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Cow, Human
  • Immunogen

    Synthetic peptide within Human Notch1 aa 2500-2600. The exact sequence is proprietary.

  • Positive control
    • WB: HEK-293 cell lysate. Mouse brain lysate. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.
  • General notes

    A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52627 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-P 1/100 - 1/150. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt 1/130 - 1/200.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/1000 - 1/2000. Detects a band of approximately 125 kDa.

 

ICC/IF 1/100 - 1/150.

 

  • Application notes
    Is unsuitable for IP.
  • 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
      • 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

    Images

    • Decrease of goblet cells in mice fed a high-fat diet (HFD).

      Immunohistochemistry using anti-Notch intracellular domain (NICD) ab52627 and phospho-S6 Abs.

      Mice intestines were flushed with phosphate-buffered saline (PBS) and fixed in 10% neural formalin overnight at room temperature. The paraffin-embedded specimens were cut into 5 μm sections and stained with hematoxylin and eosin (H&E) or periodic acid-Schiff (PAS)/Alcian blue. Paneth cells were stained with purple, and goblet cells blue with the PAS/Alcian blue method.

    • Verification of gene expression array data by immunohistochemical analysis of Notch 1 expression in subcutaneous tumors and lung metastases from a human melanoma (MeWo) xenograft experiment in mice.

      Immunohistochemical staining for Notch1 expression (ab52627, red) in subcutaneous tumors and lung metastases (both panels) of MeWo (Human malignant melanoma cell line) cells.

      All scale bars: 50 μm.

    • Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/2000 dilution (Purified) + Mouse brain at 10 µg

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Observed band size: 125 kDa
      why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

       

    • Immunohistochemical staining of paraffin-embedded human brain with purified ab52627 at a dilution of 1/150.

      A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    • Immunofluorescent staining of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells fixed with 4% PFA using purified ab52627 at a dilution of 1/150.

      An Alexa Fluor® 555 goat anti-rabbit was used as the secondary and the sample was stained with DAPI. An Alexa Fluor® 555 goat anti-mouse was used at a dilution of 1/500 after ab52627 as the negative control and is shown in the bottom right hand panel.

    • Flow cytometry analysis of permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (2% PFA, pink) with purified ab52627 at a 1/200 dilution, or negative control rabbit monoclonal IgG (green). The secondary antibody was FITC goat anti-rabbit.

    • Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/2000 dilution (Purified) + HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 10 µg

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Observed band size: 125 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemical staining of paraffin-embedded human brain with unpurified ab52627 at a dilution of 1/100.

      A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    • Immunofluorescent staining of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells fixed with 4% PFA using unpurified ab52627 at a dilution of 1/100.

      An Alexa Fluor® 555 goat anti-rabbit was used as the secondary and the sample was stained with DAPI. An Alexa Fluor® 555 goat anti-mouse was used at a dilution of 1/500 as the negative control and is shown in the bottom right hand panel.

    References

    This product has been referenced in:
    • Lv L  et al. MicroRNA-495 targets Notch1 to prohibit cell proliferation and invasion in oral squamous cell carcinoma. Mol Med Rep 19:693-702 (2019). Read more (PubMed: 30387817) »
    • Li S  et al. Notch1 inhibition enhances DNA damage induced by cisplatin in cervical cancer. Exp Cell Res 376:27-38 (2019). Read more (PubMed: 30690027) »
    See all 78 Publications for this product

    Customer reviews and Q&As

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    1-10 of 11 Abreviews

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 100X Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% tween 20
    Specification
    Liver
    Blocking step
    sea block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde

    Mr. David Ivancic

    Verified customer

    Submitted Jun 16 2019

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Balb/C liver section)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH 6
    Permeabilization
    No
    Specification
    Balb/C liver section
    Blocking step
    Innovex Background Buster as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 20°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 26 2018

    Application
    Western blot
    Sample
    Human Cell lysate - other (Lysate of cytosol from MDA468)
    Gel Running Conditions
    Reduced Denaturing (8% TG gel)
    Loading amount
    50000 cells
    Specification
    Lysate of cytosol from MDA468
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted Feb 02 2017

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (CT26-WT tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate buffer pH6
    Permeabilization
    No
    Specification
    CT26-WT tumor
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jun 07 2016

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

    Ms. Victoria Lladó

    Verified customer

    Submitted Oct 16 2015

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Airway Epithelial cells)
    Gel Running Conditions
    Reduced Denaturing (4-15%)
    Loading amount
    30 µg
    Specification
    Airway Epithelial cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

    Abcam user community

    Verified customer

    Submitted Aug 26 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Embryonic kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris pH=9
    Permeabilization
    No
    Specification
    Embryonic kidney
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jul 30 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH 9.0
    Sample
    Mouse Tissue sections (Testis)
    Specification
    Testis
    Permeabilization
    Yes - Triton
    Fixative
    Formaldehyde

    Dr. Daniel Murta

    Verified customer

    Submitted Jan 23 2014

    Application
    Western blot
    Loading amount
    100 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (ovarian cancer cell lines)
    Specification
    ovarian cancer cell lines
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Aug 01 2013

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Cow Tissue sections (tongue)
    Specification
    tongue
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer
    Permeabilization
    Yes - Triton
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C

    Dr. Daniel Murta

    Verified customer

    Submitted May 15 2013

    1-10 of 11 Abreviews

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