Detection of Human, Mouse, and Rat Nicalin by Western Blot. Western blot shows lysates of HT‑29 human colon adenocarcinoma cell line, CEM human T-lymphoblastoid cell line, Huh‑7 human hepatoma cell line, Neuro‑2A mouse neuroblastoma cell line, and NR8383 rat alveolar macrophage cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human/|
Mouse/Rat Nicalin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7686) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Nicalin at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Nicalin (NICAstrin-Like proteIN) is a 60-62 kDa member of the nicastrin family of proteins. It is an ER-embedded transmembrane protein that is found in skeletal and cardiac muscle, hepatocytes and neurons. Nicalin interacts with 22 kDa TMEM147 and 130 kDa NOMO1 to form a 200-220 kDa complex that antagonizes Nodal signaling. Mature human nicalin is a 521 amino acid (aa) type I transmembrane glycoprotein. It contains a 480 aa luminal/extracellular region (aa 43-522) plus a 20 aa cytoplasmic domain (aa 544-563). The luminal region possesses one likely inactive M28 Zn-peptidase-like domain (aa 130-429). Four potential isoform variants are reported. One shows an alternative start site at Met75, a second contains a seven aa insertion after Gln543, a third contains a 102 aa substitution for aa 162-563, and a fourth shows a deletion of Gln446. Over aa 1-522, human nicalin shares 95% aa sequence identity with mouse nicalin.