biostudies-literatureMesa RANCATS NIH HHSNIAMS NIH HHS1211https://www.ebi.ac.uk/biostudies/studies/S-EPMC7194342biostudies-literatureUnknown8Nicastrin (NCSTN) is a transmembrane glycoprotein that is part of the gamma-secretase complex. Gamma-secretase is a protease complex that cleaves type-I single-pass transmembrane proteins. There are many potential substrates for this complex, including NOTCH receptors and amyloid precursor proteins (APP). There are a number of commercial antibodies to nicastrin, but they do not agree on expected peptide size. We confirmed the specificity of a C-terminal binding rabbit anti-human antibody from Sigma-Aldrich (#N1660) using wildtype HEK293 cells and HEK293 cells deleted for nicastrin. The wildtype cells showed a prominent band at approximately 110 kDa. We confirmed this larger than expected sized was due to glycosylation by treating the lysate with peptide-N-glycosidase F (PNGase F), which reF1000ResearchValidation of a commercial antibody to detect endogenous human nicastrin by immunoblot.PMC7194342P30 AR073752UL1 TR000448Mesa RARoberson EDOfalseValidation of a commercial antibody to detect endogenous human nicastrin by immunoblot.Nicastrin (NCSTN) is a transmembrane glycoprotein that is part of the gamma-secretase complex. Gamma-secretase is a protease complex that cleaves type-I single-pass transmembrane proteins. There are many potential substrates for this complex, including NOTCH receptors and amyloid precursor proteins (APP). There are a number of commercial antibodies to nicastrin, but they do not agree on expected peptide size. We confirmed the specificity of a C-terminal binding rabbit anti-human antibody from Sigma-Aldrich (#N1660) using wildtype HEK293 cells and HEK293 cells deleted for nicastrin. The wildtype cells showed a prominent band at approximately 110 kDa. We confirmed this larger than expected sized was due to glycosylation by treating the lysate with peptide-N-glycosidase F (PNGase F), which re2019-01-01T00:00:00Z20192025-04-19T17:43:50.159Z2020-05-22T20:12:37ZS-EPMC71943423239918010.12688/f1000research.19803.2