Rat ROBO1 (also DUTT1) is a 170-200 kDa member of the four molecule ROBO family of guidance molecules (1-3). The term ROBO derives from round-about, a description of the circuitous pathway axons take in the absence of a functional ROBO gene (3, 4). Rat ROBO1 is a type I transmembrane (TM) glycoprotein that is synthesized as a 1651 amino acid (aa) precursor. It contains an 18 aa signal sequence, an 879 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 733 aa cytoplasmic region (5, 6). The ECD contains five C2-type Ig-like domains (aa 68-541) and three fibronectin (FN) type III domains (aa 561-864). The cytoplasmic region contains multiple 15-20 aa long CC (conserved cytoplasmic) motifs (C0-C3) (7, 8). Rat ROBO1 is likely to have at least one isoform. Based on the human and mouse gene, rat ROBO1 will utilize an alternate start site, creating an A (long) and B (short) isoform. The difference is the presence of a 32 aa extension at the N-terminus of the mature molecule (occurs in the A form) (9-12). Based on ROBO3 studies, this extension will impact the ability of ROBO1 to bind Slit (10). Rat ROBO1 ECD is 98% and 96% aa identical to the ECD in mouse and human ROBO1, respectively. ROBO1 serves as a repulsing molecule for axons that cross the midline. Initially, ROBO3 allows outgrowing axons to traverse the midline/floorplate. Once crossed, axons express ROBO1 which deflects neurites attempting to recross to the ipsilateral side (13). The chemorepulsant activity of ROBO1 is dependent on ROBO1 binding to SLIT1/2. Inhibition of ROBO1 is likely due to ROBO1-ROBO3 heterophilic binding (10, 13-15).
Key Product Details
Species Reactivity
Rat
Applications
Western Blot, ELISA Detection (Matched Antibody Pair)
Label
Biotin
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant rat ROBO1
Lys19-Ile560
Accession # O55005
Lys19-Ile560
Accession # O55005
Specificity
Detects rat ROBO1 in ELISAs and Western blots. In sandwich immunoassays, less than 0.5% cross-reactivity with recombinant human ROBO3 and recombinant mouse ROBO3 is observed.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Scientific Data Images for Rat ROBO1 Antibody [Biotin]
Rat ROBO1 ELISA Standard Curve
Recombinant Rat ROBO1 Fc Chimera (Catalog # 1749-RB) was serially diluted and captured by Mouse Anti-Rat ROBO1 Monoclonal Antibody (Catalog # MAB17491) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Rat ROBO1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1749) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)Applications for Rat ROBO1 Antibody [Biotin]
Application
Recommended Usage
Western Blot
0.1 µg/mL
Sample: Recombinant Rat ROBO1 Fc Chimera (Catalog # 1749-RB)
Sample: Recombinant Rat ROBO1 Fc Chimera (Catalog # 1749-RB)
Rat ROBO1 Sandwich Immunoassay
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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.
Calculators
Background: ROBO1
References
- Guthrie, S. (2001) Curr. Biol. 11:R300.
- Guthrie, S. (2004) Curr. Biol. 14:R632.
- Park, K.W. et al. (2003) Dev. Biol. 261:251.
- Seeger, M. et al. (1993) Neuron 10:409.
- Kidd, T. et al. (1998) Cell 92:205.
- Marillat, V. et al. (2002) J. Comp. Neurol. 442:130.
- Bashaw, G.J. et al. (2000) Cell 101:703.
- Jen, J.C. et al. (2004) Science 304:1509.
- Clark, K. et al. (2002) FEBS Lett. 523:12.
- Camurri, L. et al. (2005) Mol. Cell. Neurosci. 30:485.
- Sundaresan, V. et al. (1998) Mol. Cell. Neurosci. 11:29.
- GenBank Accession # O55005.
- Sabatier, C. et al. (2004) Cell 117:157.
- Liu, Z. et al. (2004) Mol. Cell. Neurosci. 26:232.
- Brose, K. et al. (1999) Cell 96:795.
Long Name
Roundabout Homolog 1
Alternate Names
DUTT1
Gene Symbol
ROBO1
UniProt
Additional ROBO1 Products
Product Documents for Rat ROBO1 Antibody [Biotin]
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Rat ROBO1 Antibody [Biotin]
For research use only
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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