Western blot analysis of total cell lysates from HepG2 and LNCaP cells, using Jagged1 (1C4) Rabbit mAb.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 263
Jagged1 (1C4) Rabbit mAb detects endogenous level of total Jagged1 protein. It does not cross-react with Jagged2.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly867 (extracellular region) of human Jagged1.
Notch signaling is activated upon engagement of the Notch receptor with its ligands, the DSL (Delta, Serrate, Lag2) proteins of single-pass type I membrane proteins. The DSL proteins contain multiple EGF-like repeats and a DSL domain that is required for binding to Notch (1,2). Five DSL proteins have been identified in mammals: Jagged1, Jagged2, Delta-like (DLL) 1, 3 and 4 (3). Ligand binding to the Notch receptor results in two sequential proteolytic cleavages of the receptor by the ADAM protease and the γ-secretase complex. The intracellular domain of Notch is released and then translocates to the nucleus where it activates transcription. Notch ligands may also be processed in a way similar to Notch, suggesting a bi-directional signaling through receptor-ligand interactions (4-6).
Mutation in Jagged1 is associated with Alagille syndrome, an autosomal dominant disorder characterized by abnormal development of liver, heart, skeleton, eye, and face (7, 8) and Tetralogy of Fallot (ToF), a common form of complex congenital heart disease (9). Jagged1 expression is associated with prostate cancer metastasis and recurrence (10).
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