Description

The 16F11 monoclonal antibody specifically binds to the extracellular domain of Notch2. This Type 1 transmembrane protein is a member of the Notch family that includes Notch1-4. Notch ligands include membrane-bound Jagged1, Jagged2 and Delta-like1, 3 and 4. Notch receptors mediate juxtacrine signals between adjacent cells. Upon ligand binding, Notch2 undergoes proteolytic cleavage that results in the release of the Notch intracellular domain, NICD. NICD translocates to the nucleus where it forms a transcriptional activator complex with the RBP-J DNA binding factor and other transcriptional cofactors. These multimeric complexes regulate the expression of multiple genes including those that orchestrate many facets of embryonic development and the subsequent functioning of cells within multiple organ systems. Notch2 is expressed on bone marrow cells, DN1 and DN2 thymocytes, B cells and activated T cells. Notch2 plays multiple roles in the immune system including the development of marginal zone B cells and the regulation of granzyme B expression by CD8+ T cells. In altered forms, Notch2 has been implicated in cellular transformation or progression in various neoplasms.

Application Notes

The antibody was conjugated to an oligonucleotide that contains an antibody clone-specific barcode (ABC) flanked by a poly-A tail on the 3" end and a PCR handle (PCR primer binding site) on the 5" end.The ABC for this antibody was designed to be used with other BD AbSeq oligonucleotides conjugated to other antibodies. All AbSeq ABC sequences were selected in silico to be unique from human and mouse genomes, have low predicted secondary structure, and have high Hamming distance within the BD AbSeq portfolio, to allow for sequencing error correction and unique mapping. The poly-A tail of the oligonucleotide allows the ABC to be captured by the BD Rhapsody™ system. The 5" PCR handle allows for efficient sequencing library generation for Illumina sequencing platforms.

NOTE:The BD Rhapsody Single-Cell Analysis System must be used with the BD Rhapsody Express Instrument.

Format

Suggested Companion Products

Preparation and Storage

Store undiluted at 4°C and protected from prolonged exposure to light.Do not freeze.The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography and conjugated to BD AbSeq oligonucleotide under optimal conditions.

Product Notices

1. This reagent has been pre-diluted for use at the recommended volume per test. Typical use is 2 µl for 1 × 10^6 cells in a 200-µl staining reaction.
2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity.However, verification testing has not been performed on all conjugate lots.
5. Illumina is a trademark of Illumina, Inc.
6. This product is covered by one or more of the following patents: US 8,835,358; US 9,290,808; US 9,290,809; US 9,315,857; US 9,567,645; US 9,567,646; US 9,598,736; US 9,708,659; and US 9,816,137.This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
8. Please refer to bd.com/genomics-resources for technical protocols.