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.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
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.
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