Notch 1 (NOTCH1) (Cleaved N-Term) antibody

Antigen: Notch 1 (NOTCH1)

Synonyms: hN1, TAN1, Mis6, Tan1, lin-12, 9930111A19Rik, NOTCH, NOTCH1, notch1, hn1, tan1, notch, xotch, xnotch, notch-1, xnotch1

Epitope: Cleaved N-Term

Clonality: Polyclonal

Host: Rabbit

Reactivity: Mouse (Murine), Human, Rat (Rattus)

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN104897

Additional Information

Alternative name: NOTCH 1 (Cleaved NT)

Swiss-Prot: P46531

Immunogen: This whole rabbit serum was prepared by repeated immunizations with a synthetic peptide corresponding to amino acid residues of human Notch 1 located near the N-terminal sequence of the cleaved N intracellular domain (NICD).

Format: Liquid (sterile filtered)

Description: Notch is synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase (S1 cleavage) in the trans-golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved (S2 cleavage) by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called Notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin-dependent gamma-secretase (S3 cleavage) to release the intracellular domain (NICD) from the membrane.

Specificity: This antiserum is directed against human NOTCH 1. Based on the immunogen sequence, we expect this antibody to react as well with mouse and rat NOTCH 1 (100% sequence homology). This antibody reacts with mouse Notch constructs present in lysates of HEK 293 cells. Only the cleaved intracellular (activated) form (NICD) is detected. No reactivity is detected against mouse N2, N3 or N4. The immunogen epitope is only exposed after gamma secretase cleavage and is not accessible in the uncleaved form.

Application Details

Application Notes: Recommended Dilutions: This product was assayed by immunoblot against recombinant mouse Notch constructs in 293 cells. An 80 kDa band corresponding to Notch 1 was observed at a 1:500 dilution. This product was also assayed against the peptide immunogen in a standard capture ELISA using Peroxidase conjugated anti-Rabbit IgG (H&L) [Goat] (code ABIN101990) and ABTS (2,2-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) (code ABTS-100) as a substrate for 30 minutes at room temperature. A working dilution of 1:20,000 to 1:60,000 is suggested for this product. Optimal titers for other applications should be determined by the researcher. Note: This antibody is reactive with Notch 1 by western blotting. A strong response was also detected by ELISA against the immunizing peptide.

Concentration: 80 mg/ml (by Refractometry)

Buffer: Stabilizer: None. Preservative: 0.1% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at -20° C prior to opening. For extended storage, aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

Research Area: Receptors, Signaling, Inflammation, Immunology

Restrictions: For Research Use only

Images

Immunoblotting. Rabbit anti-Human NOTCH 1 Cleaved N Terminal was used at a 1:500 dilution to detect mouse Notch 1 by Western blot. Equivalent amounts of lysates from transiently transfected 293 cells expressing recombinant myc-tagged mouse Notch constructs were electrophoresed and transferred to membrane using standard methods. A reaction with diluted primary antibody was followed by washings, reaction with a 1:10,000 dilution of HRP conjugated Gt-a-Rabbit IgG (ABIN102010), and color development. Lane M: Mol wt markers. Lane 1 No transfection. Lane 2 N1 (mouse deleted extracellular domain)- myc. Lane 3 N1 (mouse intracellular domain)-myc. Lane 4 N2 (mouse deleted extracellular domain)-myc. Lane 5 N2 (mouse intracellular domain)-myc. Lane 6 N3 (mouse deleted extracellular domain)-myc. Lane 7 N3 (mouse intracellular domain)-myc. Lane 8 N4 (mouse deleted extracellular domain)-myc. Lane 9 N4 (mouse intracellular domain)-myc. Lane 10 N1 (mouse deleted extracellular domain)(V to G)-myc. Data contributed by Stacey Huppert, personal communication.

Publications

Publications: Solomon, Mian, Ortega-Cava et al.: "Upregulation of the let-7 microRNA with precocious development in lin-12/Notch hypermorphic Caenorhabditis elegans mutants." in: Developmental biology, Vol. 316, Issue 2, pp. 191-9, 2008 (PubMed).

Gupta, Li, Abedin et al.: "Effect of Notch activation on the regenerative response to acute renal failure." in: American journal of physiology. Renal physiology, 2009 (PubMed).