Specific cells in the midline separate the left and right halves of the central nervous system and play many roles in guiding the growth cone behavior. In vertebrate spinal cord, insect abdominal nerves and nematodes, midline cells produce induced cues, such as nectins and slits, respectively, as attractants and repellents. respectively. These cells can serve as gatekeepers to prevent axons from passing through the midline and to guide the growth cone in response to the switching of lead cues beyond the crossing. One such gatherer, Robo, is an axon guidance receptor that defines a new subfamily of proteins from fruit flies to mammalian conserved Ig superfamily proteins. Robo acts as a receptor for the repellent Slit and functions in a cell-autonomous manner Non-intersecting axons express high levels of Robo and cross axons to express low levels of Robo and then reach midline and high levels of crossover. Robo1 and Robo2 are two human homologs of Drosophila megalopolis. Robo1 is also homologous to the C. elegans gene sax3, while Robo2 is homologous to zebrafish genes.
Background References
1. Bianchi G et al. ROBO1 Promotes Homing, Dissemination, and Survival of Multiple Myeloma within the Bone Marrow Microenvironment. Blood Cancer Discov. 2021 Jul
2. Xie J et al. Slit2/Robo1 Mitigates DSS-induced Ulcerative Colitis by Activating Autophagy in Intestinal Stem Cell. Int J Biol Sci. 2020 Apr
Western blot analysis of ROBO1 on different lysates with Mouse anti-ROBO1 antibody (HA601056) at 1/1,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane) Lane 2: Rat brain tissue lysate (40 µg/Lane)
Predicted band size: 181 kDa Observed band size: 181 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601056) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human breast tissue with Mouse anti-ROBO1 antibody (HA601056) at 1/600 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601056) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Mouse anti-ROBO1 antibody (HA601056) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601056) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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