Anti-Notch 1 Antibody, NT

Notch is a family of single-pass transmembrane receptor proteins that are synthesized in the endoplasmic reticulum as an inactive form and are proteolytically cleaved on an extracellular site by a furin-like convertase (S1 cleavage) in the trans-golgi network after the recognition of the RQRR sequence before it reaches the plasma membrane as heterodimers to yield an active, ligand-accessible form. The Notch family is comprised of 4 members (1-4) whose ligands include the Delta and Jagged family of ligands. These ligands cause proteolysis of Notch to liberate the intracellular domain. Cleavage results in a C-terminal fragment N(TN) 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 between gly1743 and val1744 (S3 cleavage) to release the intracellular domain (NICD) from the membrane. That domain translocates to the nucleus and initiates transcription events by binding the DNA binding protein CSL. The notch family members are involved cell differentiation and development.

~80 kDa

Synthetic peptide from the N-terminal sequence of the cleaved N intracellular domain (NICD) human Notch 1.

Epitope: Cleaved N-Terminus

Anti-Notch 1 Antibody, N-Terminus is a Rabbit Polyclonal Antibody for detection of Notch 1 also known as Neurogenic locus notch homolog protein 1 precursor & has been validated in ELISA, WB, IHC(P).

ELISA:
A previous lot of this antibody was diluted to 1:20,000-1:60,000 and used in a standard sandwich ELISA assay against the peptide immunogen.

Immunohistochemistry(paraffin):
Representative testing from a previous lot.
Optimal Staining of Notch-1 With Citrate pH 6.0 Epitope Retrieval: Uterine Cancer

ELISA: A previous lot of this antibody was diluted to 1:20,000-1:60,000 and used in a standard sandwich ELISA assay against the peptide immunogen.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Transcription Factors

Notch 1, cleaved N terminal. Only the cleaved intracellular (activated) form is detected. No reactivity is detected against mouse N2, N3 or N4. This epitope is exposed only after gamma secretase cleavage and is not accessible in the uncleaved form.

Other species have not been tested.

Rabbit polyclonal IgG serum in buffer containing 0.02 M Potassium Phosphate, 0.15 M NaCl, pH 7.2, containing 0.1% sodium azide.

Stable for 6 months at -20°C in undiluted aliquots from date of receipt. Avoid repeated freeze/thaw cycles.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Control
Jurkate Cell Lysates. In fetal tissues, use spleen, brain stem and lung. In adult tissues use mainly lymphoid tissues.

Routinely evaluated by Western Blot on Jurkat lysates.

Western Blot Analysis: 1:500 dilution of this lot detected cleaved NOTCH1 on 10 μg of Jurkat lysates.

Replaces: 04-1046

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