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).
Use a concentration of 1 µg/ml. Detects a band of approximately 210 kDa (predicted molecular weight: 210 kDa).
Use a concentration of 1 µg/ml.
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.
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.
Belongs to the NOTCH family. Contains 5 ANK repeats. Contains 28 EGF-like domains. Contains 3 LNR (Lin/Notch) repeats.
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.
Cell membrane and Nucleus. Following proteolytical processing NICD is translocated to the nucleus.
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.
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
Torii M et al. Analysis of the microvascular morphology and hemodynamics of breast cancer in mice using SPring-8 synchrotron radiation microangiography. J Synchrotron Radiat24:1039-1047 (2017).
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