Anti-NUMB

Numb is a 75kD member of a family of cytoplasmic adaptor proteins termed CLASPS clathrin-associated sorting proteins). These link transmembrane cargo proteins to the endocytic machinery. Numb is normally associated with the basolateral surface of polarized cells and has been shown to regulate cell fate by modulating activity of the Notch pathway. Human Numb is 651 amino acids (aa) in length. It contains one phosphotyrosine-binding domain (PTB, aa 24-172) and one proline-rich region (PRR, aa 366-413). There is a potential for multiple alternate splice forms that involve all regions of the molecule. Over aa 356-592, human Numb shares 88% and 90% aa identity with mouse and canine Numb, respectively. The hNUMB gene encodes 4 different isoforms, and the immunogen used contains sequence common to all 4 isoforms. , , Applications: , Suitable for use in Western blot, Immunocytochemistry, and Direct ELISA. Other applications have not been tested. , , Recommended Dilutions: , Western blot: 1.0 ug/ml , Immunocytochemistry: 10ug/ml Fix cwlls with PBS containing 4% paraformaldehyde for 20 minutes at RT and block with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at RT. After blocking, incubate cells with diluted primary antibody for 3 hours at RT. Follow by NorthernLightsTM 557 donkey anti-sheep lgG at RT for 1 hour. Between each step, wash cells with PBS containing 0.1% BSA. , Direct ELISA: 0.5-1.0ug/ml. The detection limit is ~0.1 ng/well. , Optimal dilutions to be determined by the researcher. , , Storage and Stability:, , Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile 40-50% glycerol, aliquot and store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.