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Rat ROBO1 Biotinylated Antibody

Catalog #: BAF1749 Datasheet / COA / SDS
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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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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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