RecombinantRabMAb

Recombinant Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18049] to NOTCH4 - BSA and Azide free
  • Suitable for: Flow Cyt, IP, ICC/IF, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free
    See all NOTCH4 primary antibodies

  • Description

    Rabbit monoclonal [EPR18049] to NOTCH4 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, IP, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment aa 1100-1300. The exact sequence is proprietary.
    Database link: Q99466

  • Positive control

    • WB: Human NOTCH4 fragment recombinant protein; Human fetal lung, placenta, fetal brain, fetal heart and fetal kidney lysates; HeLa, HepG2, A549, Jurkat, C6, RAW 264.7, PC-12, NIH/3T3 and F9 whole cell lysates; Mouse brain, heart, spleen and placenta lysates; Rat brain and heart lysates. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: HeLa and NIH/3T3 cells. IP: HeLa whole cell lysate.

  • General notes

    Ab222400 is the carrier-free version of ab184742. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab222400 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

Applications

Our Abpromise guarantee covers the use of ab222400 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
Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 150 kDa (predicted molecular weight: 210 kDa).

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 regulate branching morphogenesis in the developing vascular system.

  • Tissue specificity

    Highly expressed in the heart, moderately in the lung and placenta and at low levels in the liver, skeletal muscle, kidney, pancreas, spleen, lymph node, thymus, bone marrow and fetal liver. No expression was seen in adult brain or peripheral blood leukocytes.

  • Sequence similarities

    Belongs to the NOTCH family.
    Contains 5 ANK repeats.
    Contains 28 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.

  • Cellular localization

    Cell membrane and Nucleus. Following proteolytical processing NICD is translocated to the nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • FLJ16302 antibody
      • hNotch 4 antibody
      • hNotch4 antibody
      • INT3 antibody
      • Neurogenic locus notch homolog protein 4 antibody
      • NOTC4_HUMAN antibody
      • Notch 4 antibody
      • Notch 4 intracellular domain antibody
      • Notch homolog 4 antibody
      • Notch4 antibody
      • NOTCH4 protein antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)
      Immunocytochemistry/ Immunofluorescence - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling NOTCH4 with ab184742 at 1/200 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing cytoplasmic staining on NIH/3T3 cell line.

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184742).

    • Immunocytochemistry/ Immunofluorescence - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)
      Immunocytochemistry/ Immunofluorescence - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling NOTCH4 with ab184742 at 1/200 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing cytoplasmic staining on HeLa cell line.

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184742).

    • Flow Cytometry - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)
      Flow Cytometry - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)

      Flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling NOTCH4 with ab184742 at 1/200 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184742).

    • Flow Cytometry - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)
      Flow Cytometry - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)

      Flow cytometric analysis of 2% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling NOTCH4 with ab184742 at 1/200 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A]-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184742).

    • Immunoprecipitation - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)
      Immunoprecipitation - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)

      NOTCH4 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab184742 at 1/60 dilution.

      Western blot was performed from the immunoprecipitate using ab184742 at 1/5000 dilution.

      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.

      Lane 1: HeLa whole cell lysate, 10µg (Input).

      Lane 2: ab184742 IP in HeLa whole cell lysate.

      Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab184742 in HeLa whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 1 second.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184742).

    • Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)
      Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)

    References (1)

    Publishing research using ab222400? Please let us know so that we can cite the reference in this datasheet.

    ab222400 has been referenced in 1 publication.

    • Hu J  et al. Notch1/2/3/4 are prognostic biomarker and correlated with immune infiltrates in gastric cancer. Aging (Albany NY) 12:2595-2609 (2020). PubMed: 32028262

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