Overview

  • Product name

  • 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.
  • Species reactivity


    Predicted to work with: Human
  • 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

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 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.

  • 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.
  • 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.
  • 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

This product has been referenced in:

  • 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). Read more (PubMed: 30975104) »
  • Davis RB  et al. Notch signaling pathway is a potential therapeutic target for extracranial vascular malformations. Sci Rep 8:17987 (2018). Read more (PubMed: 30573741) »
See all 7 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Kidney, bowman's capsule Cells)
Specification
Kidney, bowman's capsule Cells
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% Triton X 100
Blocking step
BSA+FBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Aug 05 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (skeletal muscle)
Specification
skeletal muscle
Fixative
PFA 1%
Permeabilization
Yes - PBS-Triton X-100 0.3%
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 11 2010

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

Sign up