Notch1, Extracellular Domain (Notch Homolog Protein 1, hN1, Translocation-associated Notch Protein TAN-1)

The Notch signaling pathway, first identified in Drosophila, regulates the generation of differentiated cells from a population of undifferentiated progenitors. The Notch 1 protein is a single pass transmembrane receptor activated through the Delta (Dl) and Serrate (Ser) ligands in Drosophila (and their mammalian counterparts Delta-like-3 and Jagged 1 and 2). Activation results in the proteolytic cleavage of the cytoplasmic portion of Notch (120kD) which localizes to the nucleus to interact with DNA binding proteins (RB-Jk/KBF2/CBF1). The interaction of Notch with these proteins converts them from transcriptional repressors to activators, thus affecting the expression of a variety of target genes. During maturation, the Notch molecule is cleaved by furin-like convertase at the extracellular domain. Upon binding to a ligand such as Delta1, or upon extracellular calcium depletion, the C-terminal Notch 1 fragment dissociates from its N-terminal counterpart and is further cleaved between Gly1743 and Val1744. The resulting activated cytosolic fragment translocates to the nucleus and activates Notch-related transcription.\n\nApplications:\nSuitable for use in Immunoblot anaylsis. Other applications have not been tested.\n\nRecommended Dilution:\nImmunoblot Analysis: 1ug/ml detects Notch 1 in a RIPA lysate of L/N17 rat Notch 1 transfected fibroblast cells. L/N17 transfected Notch1 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Notch1 (1ug/ml). Proteins visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability: \nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.\n\nSpecial Note:\nThis product was previously supplied as 200ul in the same buffer with dilution instructions of 1:1000, yielding 100 Western Blots/vial using 10ml buffer : 10ul antibody.\nThe same number of tests are supplied when using 1ug/ml, ie, 100ug product will yield 100 Western Blots.

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SPECIFICATIONS

Catalog Number

N5375-10

Size

100ug

Applications

WB

Hosts

Rabbit

Reactivities

Rat

Form

Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide before the addition of glycerol to 30%.

P Type

Pab

Purity

Purified by Protein A affinity chromatography.

Isotype

IgG

References

1. Blaumueller, C.M., et al., Cell 90: 281-291, 1997 2. Garces, C., et al., J. Biol. Chem. 272: 29729-29734, 1997 3. Jarriault, S., et al., Mol. Cell. Biol. 18: 7423-431, 1998 4. Oswald, F., et al., Mol. Cell. Biol. 18: 2077-2088, 1998 5. Struhl, G., et al., Cell 93: 649-660, 1998 6. Weinmaster, G., Cur. Op. in Gen. & Dev. 8: 436-442, 1998.

Additional Info

Recognizes rat Notch 1 at 260kD. Species crossreactivity: Rat