ANTI-NOTCH 2 ANTIBODY NT

General description

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.

Specificity

Notch 2, cleaved N terminal. This epitope is exposed only after gamma secretase cleavage and is not accessible in the uncleaved form.

Other species have not been tested.

Immunogen

Epitope: Cleaved N-terminus

Synthetic peptide from the N-terminal sequence of the cleaved N intracellular domain (NICD) human Notch 2.

Application

Anti-Notch 2 Antibody, N-Terminus detects level of Notch 2 & has been published & validated for use in ELISA, WB, IH(P).

ELISA:
1:30,000-1:90,000 dilution from a previous lot was used in a standard sandwich ELISA assay against the peptide immunogen.
Optimal working dilutions must be determined by the end user.

Immunohistochemistry(paraffin):
Representative testing from a previous lot.
Optimal Staining of NOTCH-2 Polyclonal Antibody: Squamous Cell Carcinoma

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Transcription Factors

Quality

Routinely evaluated by Western Blot on NIH/3T3 lysates.

Western Blot Analysis: 1:500 dilution of this lot detected cleaved NOTCH2 on 10 μg of NIH/3T3 lysates.

Target description

~72 kDa

Linkage

Replaces: PART AB5713

Physical form

Rabbit polyclonal IgG serum in buffer containing 0.02 M Potassium phosphate buffer with 0.15 M NaCl and 0.1% sodium azide

Storage and Stability

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.

Analysis Note

Control
NIH/3T3 Cell Lysate

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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