Notch4 is a 250kD heterodimeric membrane protein with a C-terminal transmembrane fragment covalently linked to an N-terminal extracellular fragment.
It functions as a receptor for transmembrane ligands such as the Jagged and Delta proteins.
Upon ligand binding, it undergoes sequential proteolytic processing to release the intracellular domain (NICD), which is translocated to the nucleus where it activates target genes.
In the region used as immunogen, human and mouse Notch4 share 78% amino acid sequence homology.
Notch signaling plays a key role in the normal development of many tissues and cell types, through diverse effects on differentiation, survival, and/or proliferation that are highly dependent on signal strength and cellular context.
Members of the Notch gene family encode transmembrane receptors that are critical for various cell fate decisions.
Notch family members share structural characteristics including an extracellular domain consisting of multiple epidermal growth factor-like (EGF) repeats, and an intracellular domain consisting of multiple, different domain types.
Multiple human notch proteins (Notch1, Notch2, Notch3 and Notch4) have been identified and they function as a receptors for membrane bound ligands.
Notch signaling is also linked to tumorigenesis as first demonstrated by the identification of a recurrent t(7;9)(q34;q34.3) chromosomal translocation involving the human Notch1 gene that is found in a small subset of human pre-T-cell acute lymphoblastic leukemias (T-ALL).
Since this discovery, aberrant Notch signaling has been suggested to be involved in a wide variety of human neoplasms.
Notch4 functions as a receptor for membrane bound ligands, and may play a role in vascular, renal and hepatic development.
Notch4 may also be associated with susceptibility to schizophrenia in a small portion of cases.
Applications:Western Blot: 0.1-0.2ug/ml with the appropriate secondary reagents to detect human Notch4 intracellular domain.
The detection limit for rhNotch4 intracellular domain is approximately 5ng/lane under non-reducing and reducing conditions.
Immunocytochemistry: 10ug/ml to detect human Notch4 in serum starved HUVEC cells stimulated with Jagged 1.
Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at RT and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at RT.
After blocking, cells were incubated with diluted primary antibody overnight at 4°C followed by Rhodamine Red-coupled anti-goat IgG at RT in the dark for one hour.
Between each step, cells were washed with PBS containing 0.1% BSA.
Direct ELISA: 0.5-1.0ug/ml with the appropriate secondary reagents to detect human Notch4 intracellular domain.
The detection limit for rhNotch4 intracellular domain is approximately 0.3ng/well.
Optimal dilutions should be determined by each laboratory for each application.
Storage and Stability:Lyophilized powder may be stored at 4°C for short-term only.
Reconstitute to nominal volume by adding sterile PBS with 40-50% glycerol and store at -20°C.
Reconstituted product is stable for 12 months at -20°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.
仕様
Size：100ug
Host：goat
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by human Notch4 intracellular domain affinity chromatography.
Form：Supplied as a lyophilized powder from a 0.2um filtered solution in PBS with 5% trehalose. Reconstitute with sterile PBS, 40-50% glycerol. If 0.5ml is used, the concentration will be 0.2mg/ml.
