Notch-1 (so named for “notches” in fly wings; also TAN-1) is a 300kD member of the Notch family of glycoproteins.
It is associated with gene activation in both embryo and adult.
Human Notch-1 is a 2538 amino acid (aa) type I transmembrane glycoprotein.
It undergoes Golgi processing to generate a heterodimer composed of a 180-200kD disulfide-linked extracellular domain (aa 18-1664) and a 120kD membrane- bound segment aa 1665-2556).
Upon ligand binding, the 110kD segment undergoes two cleavages which generate an NICD (notch intracellular domain) (aa 1754-2556), a nuclear transcription factor.
One isoform shows a deletion of aa 248-288.
Applications: Suitable for use in for use in Western blot, Immunocytochemistry, Flow cytometry and Direct ELISA.
Other applications have not been tested.
Recommended Dilution:Western Blot: 0.1-0.2 ug/ml.
The detection limit for rhNotch-1 is ~ 0.5 ng/lane under non-reducing and reducing conditions.
Immunocytochemistry: 10ug/ml to detect human Notch-1 in SAOS human osteosarcoma cells.
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 for 3 hours at RT followed by NorthernLightsTM 557 donkey anti-sheep lgG at RT for 1 hour.
Between each step, cells were washed with PBS containing 0.1% BSA.
Flow cytometry: 50ug/ml and add 10il of the diluted solution to 1-2.5 x 10e5 cells in a total reaction volume not exceeding 200ul.
The binding of unlabeled antibodies may be visualized by adding 10il of a 25ug/ml stock solution of a secondary developing reagent such as anti-sheep lgG conjugated to a fluorochrome.
Direct ELISA: 0.5-1.0ug/ml with the appropriate secondary reagents to detect human Notch-1.
The detection limit is ~0.2 ng/well.
Optimal dilutions should be determined by each laboratory for each application.
Storage and Stability:Lyophilized powder may be stored at -20°C.
Reconstitute to nominal volume by adding sterile buffer.
Aliquot 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：sheep
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by affinity chromatography.
Form：Supplied as a lyophilized powder in PBS with 5% trehalose. Reconstitute with 0.5ml sterile PBS to a concentration 0.2 mg/ml.
Specificity：Recognizes human Notch-1 intracellular domain. Species sequence homology: canine 83% and mouse 89%. < 1% cross-reactivity with rhNotch-2 IC, rhNotch-3 IC, and rhNotch-4 IC.
Isotype：IgG
Calc Applications Abbrev：E FC IC WB
Calc Crossreactivity：Hu
Immunogen：E. coli-derived recombinant human rhNotch-1 (aa 2428 - 2556).
