Anti-Notch1 EGF12 T(O-Fuc)466 Monoclonal Antibody

Description    

Rabbit monoclonal antibody to human Epidermal growth factor-like repeat 12 (EGF-12) modified at threonine 466 (T466) with an O-linked fucose (O-Fuc). GS0331 binds to the O-Fucosylated version of EGF12, and also shows binding to other O-Fucosylated growth factor-like repeats 8 (T311) and EGF26 (T997).  The mAb does not detect non-fucosylated forms of these sequences. 

Further validation of this monoclonal was conducted on the EGF portion of Notch1 proteins produced in E. coli.  In vitro treatment of these proteins with glycosyltransferase POFUT1, which extends O-fucose on properly folded proteins results in a terminal fucose monosaccharide.  Further addition of the Lunatic Fringe (Lfng) glycosyltransferase in the presence of UDP-GlcNAc results in an O-Fucose containing disaccharide.  The GS0331 EGF12 mAb is able to preferentially detect reduced forms of the POFUT1 treated EGF proteins.  Treatment with PNGaseF does not inhibit detection, further verifying that the antibody is specific to an O-linked sugar rather than n-linked glycosylation (data not shown).    

Please inquire for availability of this mAb.  Production lead times may be associated with ordering this product.     

Tested applications  Direct peptide ELISA, Western blot

Target sequence      QNDAT(O-Fuc)CLDQ

Click here for a sample data sheet.