Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NOTCH4 antibody (ab33163)

Key features and details

  • Rabbit polyclonal to NOTCH4
  • Suitable for: WB, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-NOTCH4 antibody
    See all NOTCH4 primary antibodies

  • Description

    Rabbit polyclonal to NOTCH4

    This product is a fast track antibody. It has been affinity purified and shows high titre values against the immunizing peptide by ELISA. Read the terms of use »

  • Host species

    Rabbit

  • Specificity

    By sequence similarity, the Notch intracellular domain (NICD)is believed to be contained within amino acids 1467 to 2003. The peptide used for this antibody is found within this region.

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide corresponding to Human NOTCH4 aa 1800-1900 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab26877)

Properties

Applications

Fast track antibodies constitute a diverse group of products that have been released to accelerate your research, but are not yet fully characterized. They have all been affinity purified and show high titre values against the immunizing peptide (by ELISA). Fast track terms of use

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
ICC/IF
Human
WB
Human
Application Abreviews Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 210 kDa (predicted molecular weight: 210 kDa).
ICC/IF (2)
Use a concentration of 1 µg/ml.
Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 210 kDa (predicted molecular weight: 210 kDa).
ICC/IF
Use a concentration of 1 µg/ml.

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

    This Fast-Track antibody is not yet fully characterised. These images represent inconclusive preliminary data.

    • Western blot - Anti-NOTCH4 antibody (ab33163)
      Western blot - Anti-NOTCH4 antibody (ab33163)
      Anti-NOTCH4 antibody (ab33163) at 1 µg/ml + Human kidney tissue lysate - total protein (ab30203) at 20 µg

      Secondary
      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 210 kDa
      Observed band size: 43 kDa
      why is the actual band size different from the predicted?



      NOTCH4 is synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved before it reaches the plasma membrane to yield an active, ligand-accessible form. Following ligand binding, it is again cleaved to yield a membrane-associated intermediate fragment called notch extracellular truncation (NEXT). This fragment is cleaved yet another time to release the NOTCH intracellular domain (NICD) from the membrane, which translocates to the nucleus where it forms a transcriptional activation complex. The immunogen used for ab33163 is a peptide found within this NICD, and we believe it recognizes this region in western blot analysis.
    • Immunocytochemistry/ Immunofluorescence - Anti-NOTCH4 antibody (ab33163)
      Immunocytochemistry/ Immunofluorescence - Anti-NOTCH4 antibody (ab33163)
      ICC/IF image of ab33163 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab33163, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293 cells at 1µg/ml.
    • Western blot - Anti-NOTCH4 antibody (ab33163)
      Western blot - Anti-NOTCH4 antibody (ab33163)
      Anti-NOTCH4 antibody (ab33163) at 1 µg/ml + NOTCH4 overexpression lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 210 kDa
      Observed band size: 210,238 kDa why is the actual band size different from the predicted?
      Additional bands at: 52 kDa, 60 kDa. We are unsure as to the identity of these extra bands.

    References (7)

    Publishing research using ab33163? Please let us know so that we can cite the reference in this datasheet.

    ab33163 has been referenced in 7 publications.

    • McClements L  et al. FKBPL and its peptide derivatives inhibit endocrine therapy resistant cancer stem cells and breast cancer metastasis by downregulating DLL4 and Notch4. BMC Cancer 19:351 (2019). PubMed: 30975104
    • Davis RB  et al. Notch signaling pathway is a potential therapeutic target for extracranial vascular malformations. Sci Rep 8:17987 (2018). PubMed: 30573741
    • Idowu J  et al. Aberrant Regulation of Notch3 Signaling Pathway in Polycystic Kidney Disease. Sci Rep 8:3340 (2018). PubMed: 29463793
    • Martín-Gayo E  et al. Spatially restricted JAG1-Notch signaling in human thymus provides suitable DC developmental niches. J Exp Med 214:3361-3379 (2017). PubMed: 28947612
    • Torii M  et al. Analysis of the microvascular morphology and hemodynamics of breast cancer in mice using SPring-8 synchrotron radiation microangiography. J Synchrotron Radiat 24:1039-1047 (2017). PubMed: 28862627
    • Hill-Felberg S  et al. Notch receptor expression in human brain arteriovenous malformations. J Cell Mol Med 19:1986-93 (2015). WB . PubMed: 25846406
    • Sharma M  et al. Inhibition of Notch pathway attenuates the progression of human immunodeficiency virus-associated nephropathy. Am J Physiol Renal Physiol 304:F1127-36 (2013). PubMed: 23389453

    Images

    • Western blot - Anti-NOTCH4 antibody (ab33163)
      Western blot - Anti-NOTCH4 antibody (ab33163)
      Anti-NOTCH4 antibody (ab33163) at 1 µg/ml + Human kidney tissue lysate - total protein (ab30203) at 20 µg

      Secondary
      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 210 kDa
      Observed band size: 43 kDa
      why is the actual band size different from the predicted?



      NOTCH4 is synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved before it reaches the plasma membrane to yield an active, ligand-accessible form. Following ligand binding, it is again cleaved to yield a membrane-associated intermediate fragment called notch extracellular truncation (NEXT). This fragment is cleaved yet another time to release the NOTCH intracellular domain (NICD) from the membrane, which translocates to the nucleus where it forms a transcriptional activation complex. The immunogen used for ab33163 is a peptide found within this NICD, and we believe it recognizes this region in western blot analysis.
    • Immunocytochemistry/ Immunofluorescence - Anti-NOTCH4 antibody (ab33163)
      Immunocytochemistry/ Immunofluorescence - Anti-NOTCH4 antibody (ab33163)
      ICC/IF image of ab33163 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab33163, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293 cells at 1µg/ml.
    • Western blot - Anti-NOTCH4 antibody (ab33163)
      Western blot - Anti-NOTCH4 antibody (ab33163)
      Anti-NOTCH4 antibody (ab33163) at 1 µg/ml + NOTCH4 overexpression lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 210 kDa
      Observed band size: 210,238 kDa why is the actual band size different from the predicted?
      Additional bands at: 52 kDa, 60 kDa. We are unsure as to the identity of these extra bands.

    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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