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Neuroscience Neurology process Notch Pathway

Anti-Notch1 antibody [EP1238Y] (ab52627)

Price and availability

385 568 ₸

Availability

Order now and get it on Tuesday March 02, 2021

Anti-Notch1 antibody [EP1238Y] (ab52627)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1238Y] to Notch1
  • Suitable for: IHC-P, Flow Cyt, WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

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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 & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    Human
    See all applications and species data
  • Immunogen

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

  • Positive control

    • WB: HeLa, HAP1, MOLT-4, HEK-293 cell lysate. Mouse brain lysate. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.
  • General notes

    Notch 1 (encoded by the Notch 1 gene) is a member of the Notch family of receptors. When bound to a ligand, the Notch1 receptor sends signals critical for normal development of many tissues throughout the body. Notch1 signalling supports the determination and specialization of cells as well as playing a role in cell proliferation, differentiation, and apoptosis.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1238Y
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Notch Pathway
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Stem Cells
    • Signaling Pathways
    • Notch
    • Surface Molecules
    • Stem Cells
    • Hematopoietic Progenitors
    • Surface Molecules
    • Cardiovascular
    • Heart
    • Cardiogenesis
    • Transcription factors/regulators
    • Developmental Biology
    • Embryogenesis
    • Somatogenesis

Images

  • Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
    All lanes : Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : NOTCH1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.


    Lanes 1- 2: Merged signal (red and green). Green - ab52627 observed at 110 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab52627 was shown to react with Notch1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261762 (knockout cell lysate ab257006) was used. Wild-type HeLa and NOTCH1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab52627 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627) Lee et al PLoS One. 2017 Nov 6;12(11):e0187515. doi: 10.1371/journal.pone.0187515. eCollection 2017. Fig 3. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] (ab52627)

    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.

  • Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
    All lanes : Anti-Notch1 antibody [EP1238Y] (ab52627) at 1 µg/ml

    Lane 1 : Wild-type HAP1 cell lysate at 40 µg
    Lane 2 : NOTCH1 knockout HAP1 cell lysate at 40 µg
    Lane 3 : MOLT-4 cell lysate at 20 µg
    Lane 4 : HepG2 cell lysate at 20 µg

    Performed under reducing conditions.

    Observed band size: 105 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab52627 observed at 105 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab52627 was shown to react with Notch1 in wild-type HAP1 cells in western blot. Loss of signal was observed when NOTCH1 knockout sample was used. Wild-type and NOTCH1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with ab52627 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
    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.

     

  • Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627) Ernst et al PLoS One. 2018 Feb 12;13(2):e0192525. doi: 10.1371/journal.pone.0192525. eCollection 2018. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627)

    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.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627)

    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.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] (ab52627)

    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.

  • Flow Cytometry - Anti-Notch1 antibody [EP1238Y] (ab52627)
    Flow Cytometry - Anti-Notch1 antibody [EP1238Y] (ab52627)

    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)
    Anti-Notch1 antibody [EP1238Y] (ab52627)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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