Anti-NUMB Antibody, clone Ab21

Protein numb homolog (UniProt: P49757; also known as h-Numb, Protein S171) is encoded by the NUMB (also known as C14orf41) gene (Gene ID: 8650) in human. NUMB is an evolutionarily conserved endocytic protein that make the daughter cells different after asymmetric divisions by segregating to only one daughter. NUMB is a peripheral membrane protein present in cell and endosome membranes. It is required throughout embryonic neurogenesis to maintain neural progenitor cells by allowing their daughter cells to choose progenitor over neuronal cell fate. However, they are not required for the proliferation of neural progenitor cells before the onset of neurogenesis. It is also involved postnatally in the subventricular zone (SVZ) neurogenesis by regulating SVZ neuroblasts survival and ependymal wall integrity. NUMB is shown to interact with AP-2 and clathrin-coated pits via -adaptin protein. It undergoes phosphorylation at serine 276 and 295 by CAMK1, which decouples it from the recycling machinery and results in inhibition of its interaction with AP-2 and clathrin-mediated endocytosis. NUMB also functions as an oncosuppressor by inhibiting Notch signaling and stabilizing p53. This effect is dependent upon its interaction with Mdm2 that ubiquitinates p53 and commits it to proteasomal degradation. Nine isoforms of NUMB have been described that are produced by alternative splicing. (Ref.: Colaluca, IN., et al. (2018). J. Cell Biol. 217(2); 745-762; Tasoni, D., et al. (2015). J. Cell. Biol. 211(4); 845-862; Tang, H., et al. (2005). Mol. Cell. Biology 25(8); 2899-2909; Tokomitsu H., et al. (2005). J. Biol. Chem. 280(42); 35108-35118).

Clone Ab21 is a mouse monoclonal antibody that detects Protein numb homolog (NUMB). It targets an epitope within 15 amino acids from the C-terminal half.

KLH-conjugated linear peptide corresponding to 15 amino acids from the C-terminal half of human NUMB protein.

Quality Control Testing

Evaluated by Western Blotting in Neuro2a cell lysate.

Western Blotting Analysis: A 1:500 dilution of this antibody detected NUMB in Neuro2a cell lysate.

Tested Applications

Western Blotting Analysis: A 1:500 dilution from a representative lot detected NUMB in A549 and U87MG cell lysates.

Immunofluorescence Analysis: A representative lot detected NUMB in Immunofluorescence application (Colaluca, I.N., et al. (2008). Nature. 451(7174):76-80; Tosoni, D., et al. (2015). J Cell Biol. 211(4):845-62; Santoro, A., et al. (2019). Cell Rep. 26(3):624-638.e8).

Immunoprecipitation Analysis: A representative lot immunoprecipitated NUMB in Immunoprecipitation application (Colaluca, I.N., et al. (2008). Nature. 451(7174):76-80; Colaluca, I.N., et al. (2018). J Cell Biol. 217(2):745-762).

Western Blotting Analysis: A representative lot detected NUMB in Western Blotting application (Colaluca, I.N., et al. (2008). Nature. 451(7174):76-80; Tosoni, D., et al. (2015). J Cell Biol. 211(4):845-62; Tosoni, D., et al. (2017). EMBO Mol Med. 9(5):65-671; Colaluca, I.N., et al. (2018). J Cell Biol. 217(2):745-762).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommended storage: +2°C to +8°C.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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