|Detection of Human ROBO1 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and SH‑SY5Y human neuroblastoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human ROBO1 Monoclonal Antibody (Catalog # MAB7118) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for ROBO1 at approximately 220 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
ROBO1 (Roundabout-like protein 1) is a 190-230 kDa member of the ROBO/roundabout receptor family. It is expressed by commissural axons from multiple nuclei, and is also found on vascular endothelium, bronchial epithelium, and synctiotrophoblasts. It interacts with Slit and DCC to temporally regulate the migration of axonal processes. The human ROBO1 precursor is a 1651 amino acid (aa) type I transmembrane protein. It contains a 25 aa signal sequence, followed by an 872 aa extracellular region (aa 26-897) that possesses five C2-type Ig-like domains (aa 68-541) and three fibronectin type III domains (aa 561-864). ROBO1 shows multiple isoform variants. The variant used here is termed ROBO1b/DUTT1, and it possesses an 18 aa substitution for aa 1‑47, accompanied by a three aa insertion after Gln348, and a deletion of aa 939-947. ROBO1a, by contrast, possesses only the 18 aa substitution just described. A third isoform possesses the same changes as ROBO1b plus an additional deletion of aa 1013-1067, while a final variant utilizes an alternative start site at Met120. Proteolytic cleavage generates a soluble 120 kDa N-terminal fragment. Over aa 20-861, human ROBO1b shares 97% aa identity with mouse ROBO1.
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