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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | WB, ICC, EM | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABC594 |
| Description | Anti-NOTCH 3/N3ECD Antibody, clone 1E4 |
| Alternate Names |
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| Background Information | NOTCH3 is a neuronal cell fate regulating protein that initially functions as a receptor for the membrane bound ligands Jagged1, Jagged2 and Delta1. NOTCH3 gene functions operate via the intracellular domain of Notch3 (NICD) that forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus which ultimately influences cellular differentiation, proliferation and apoptotic programs. NOTCH3 is widely expressed in fetal and adult tissues. NOTCH3 protein begins as an inactive form in the endoplasmic reticulum that becomes processed to an active form in the plasma membrane. Functionally, NOTCH3 is cleaved by TACE to yield a membrane bound extracellular fragment called NEXT which is then cleaved by gamma-secretase to release the NICD intracellular peptide from the membrane, which then proceeds to be translocated to the nucleus for gene the gene activations. Mutations in NOTCH3 lead to cerebral arteriopathy with subcortical infarcts and leukoencephalopathy syndrome and myofibromatosis infantile 2 disorder, both seriously debilitating genetic diseases. Disease-causing mutations affect cysteine residues within epidermal growth factor-like repeat domains in the ectodermal domain of NOTCH3 (N3ECD). One of the main biochemical and histopathological hallmarks of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the accumulation of N3ECD at the cell surface of vascular smooth muscle cells which degenerate over the course of the disease. |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This Anti-NOTCH 3/N3ECD Antibody, clone 1E4 is validated for use in Western Blotting and Immunocytochemistry and Electron Microscopy for the detection of NOTCH 3/N3ECD. |
| Key Applications |
|
| Application Notes | Immunocytochemistry Analysis: A representative lot of this antibody was used to detect NOTCH 3 in CADASIL vascular smooth muscle cells (Tikka, S., et al. (2012) Journal of Cerebral Blood Flow & Metabolism. 1–10). Immunohistochemistry Analysis: A representative lot of this antibody was used to detect NOTCH 3 in Huamn vascular smooth muscle cells & smooth muscle cells of vessels from CADASIL brain tissue (Jouet, A., et al. (2000) Journal of Clinical Investigation. 105(5):597-605). Electron Microscopy: A representative lot of this antibody was used to detect NOTCH 3 in smooth muscle cells of vessels from CADASIL brain tissue (Jouet, A., et al. (2000) Journal of Clinical Investigation. 105(5):597-605). Immunohistochemistry Analysis: A representative lot of this antibody was used to detect NOTCH 3 in vessels of a CADASIL patient (Rouchox, M.M., et al. (2003) American Journal of Pathology. 162(1):329-342). Immunohistochemistry Analysis: A representative lot of this antibody was used to detect NOTCH 3 in Human colorectal carcinoma (Serafin, V., et al., (2011) Journal of Pathology. 224(4):448-60). |
| Biological Information | |
|---|---|
| Immunogen | His-tagged recombinant protein corresponding to Human NOTCH 3. |
| Clone | 1E4 |
| Concentration | 1 mg/mL |
| Host | Mouse |
| Specificity | This antibody detects both the full length NOTCH 3 protein and the NOTCH 3 ectodermal domain (N3ECD). |
| Isotype | IgG1κ |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein G Purified |
| UniProt Number |
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| Molecular Weight | ~260 and ~210 kDa observed This antibody detects both the full length NOTCH 3 protein (~260 kDa) and the ectodermal domain N3ECD (~210 kDa). |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in MCF-7 cell lysate. Western Blotting Analysis: 1 µg/mL of this antibody detected NOTCH 3 & the ectodermal domain (N3ECD) in 10 µg of MCF-7 cell lysate. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |