Rat ROBO1 Biotinylated Antibody

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BAF1749
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Rat ROBO1 Biotinylated Antibody Summary

Species Reactivity
Rat
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.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant rat ROBO1
Lys19-Ile560
Accession # O55005
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Label
Biotin

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Rat ROBO1 Fc Chimera (Catalog # 1749-RB)

Rat ROBO1 Sandwich Immunoassay

Recommended Concentration
Reagent
ELISA Detection (Matched Antibody Pair)
0.1-0.4 µg/mL 

Use in combination with:

Capture Reagent: Rat ROBO1 Antibody (Catalog # MAB17491)

Standard: Recombinant Rat ROBO1 Fc Chimera Protein, CF (Catalog # 1749-RB)

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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.

Background: ROBO1

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).

References
  1. Guthrie, S. (2001) Curr. Biol. 11:R300.
  2. Guthrie, S. (2004) Curr. Biol. 14:R632.
  3. Park, K.W. et al. (2003) Dev. Biol. 261:251.
  4. Seeger, M. et al. (1993) Neuron 10:409.
  5. Kidd, T. et al. (1998) Cell 92:205.
  6. Marillat, V. et al. (2002) J. Comp. Neurol. 442:130.
  7. Bashaw, G.J. et al. (2000) Cell 101:703.
  8. Jen, J.C. et al. (2004) Science 304:1509.
  9. Clark, K. et al. (2002) FEBS Lett. 523:12.
  10. Camurri, L. et al. (2005) Mol. Cell. Neurosci. 30:485.
  11. Sundaresan, V. et al. (1998) Mol. Cell. Neurosci. 11:29.
  12. GenBank Accession # O55005.
  13. Sabatier, C. et al. (2004) Cell 117:157.
  14. Liu, Z. et al. (2004) Mol. Cell. Neurosci. 26:232.
  15. Brose, K. et al. (1999) Cell 96:795.
Long Name
Roundabout Homolog 1
Entrez Gene IDs
6091 (Human); 58946 (Rat)
Alternate Names
DUTT1; DUTT1Drosophila) homolog 1; H-Robo-1; MGC131599; MGC133277; ROBO1; roundabout, axon guidance receptor, homolog 1 (Drosophila)

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