Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)

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 - Low endotoxin, Azide free
  • Suitable for: ICC/IF, IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free
    See all Notch1 primary antibodies

  • Description

    Rabbit monoclonal [EP1238Y] to Notch1 - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IHC-P, Flow Cyt, WBmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Cow

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK293, HAP1 and MOLT-4 cell lysateS. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.

  • General notes

    ab246693 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
    Western blot - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
    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.

    Predicted band size: 272 kDa
    Observed band size: 105 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab52627).

    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 3pc 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627). 

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

  • Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
    Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627). 

  • Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
    Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627). 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627). 

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627). 

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627). 

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

  • Flow Cytometry - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
    Flow Cytometry - Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627). 

  • Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
    Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)

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