Recombinant Human ROBO1/DUTT1 Fc Chimera Protein, CF

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human ROBO1/DUTT1 Fc Chimera Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. When recombinant human Slit3 is coated at 250 ng/mL, Recombinant Human ROBO1 Fc Chimera binds with a typical ED₅₀ of
  100-600 ng/mL.
• Source: Mouse myeloma cell line, NS0-derived human ROBO1 protein
  

  Human ROBO1/DUTT1 (Ser20-Pro858) Accession # NP_598334.1	IEGRMD	Human IgG1 (Pro100-Lys330)
  N-terminus		C-terminus

• Accession #: NP_598334.1
• N-terminal Sequence: Ser20
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Recombinant Proteins
• Gene: ROBO1
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 119 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human ROBO1/DUTT1 Fc Chimera Protein, CF

• DUTT1
• DUTT1Drosophila) homolog 1
• H-Robo-1
• MGC131599
• MGC133277
• ROBO1
• roundabout, axon guidance receptor, homolog 1 (Drosophila)

Background

ROBO1 is a 170-200 kDa member of the ROBO family of guidance molecules. It serves as a repulsing molecule that prevents axons from crossing the midline in the developing central nervous system (CNS) (1). Mature human ROBO1 consists of an 872 amino acid (aa) extracellular domain (ECD), a 21 aa transmembrane segment, and a 733 aa cytoplasmic region (2). The ECD contains five tandem Ig-like domains followed by three tandem fibronectin type III domains. Usage of an alternative promoter generates a shorter isoform (ROBO1b or DUTT1) that has an 18 aa substitution for the N-terminal 57 residues (2, 3). Within the ECD, human ROBO1 shares 97% aa identity with mouse and rat ROBO1. A 120 kDa soluble fragment of the ECD can be shed by juxtamembrane proteolytic cleavage (4). ROBO1 is expressed on pioneer axons in the neural tube and proliferative zones of the cortical plate (5-8). It is restricted to axonal growth cones that do not cross the midline and to midline-crossing axons only after they cross (2). ROBO1b shows even wider expression in the developing mouse CNS and is the predominant isoform in spinal cord commissural axons (5). ROBO1 plays an important role in axon pathfinding and tract development (6, 9, 10) as well as in neuronal proliferation and dendrite branching (7, 8). ROBO1 binds to Slit1 and Slit2, leading to growth cone collapse and axonal branching (7, 11). It also regulates Netrin-1 and Semaphorin 3A and 3F responsiveness through its ability to form in cis complexes with Neuropilin-1, Neuropilin-2, Plexin A1, DCC, and FLRT3 (10, 12, 13). ROBO1 is additionally expressed in developing ductal epithelia where its binding to Slit2 restricts branching morphogenesis (14). ROBO1-Slit2 interactions can also suppress tumorigenesis (15), enhance VEGF-induced angiogenesis (16), and enhance CXCL12 induced T cell chemotaxis by associating with CXCR4 (17).

1. Dudanova, I and R. Klein (2013) Trends Neurosci. 36:295.
2. Kidd, T. et al. (1998) Cell 92:205.
3. Sundaresan, V. et al. (1998) Mol. Cell. Neurosci. 11:29.
4. Seki, M. et al. (2010) FEBS Lett. 584:2909.
5. Nural, H.F. et al. (2007) Gene Expr. Patterns 7:837.
6. Wang, G. et al. (2013) Exp. Cell Res. 319:1083.
7. Whitford, K.L. et al. (2002) Neuron 33:47.
8. Yeh, M.L. et al. (2014) J. Neurosci. 34:5717.
9. Andrews, W. et al. (2006) Development 133:2243.
10. Hernandez-Miranda, L.R. et al. (2011) J. Neurosci. 31:6174.
11. Liu, Z. et al. (2004) Mol. Cell. Neurosci. 26:232.
12. Stein, E. and M. Tessier-Lavigne (2001) Science 291:1928.
13. Leyva-Diaz, E. et al. (2014) Curr. Biol. 24:494.
14. Macias, H. et al. (2011) Dev. Cell 20:827.
15. Chang, P.H. et al. (2012) Cancer Res. 72:4652.
16. Rama, N. et al. (2015) Nat. Med. 21:483.
17. Prasad, A. et al. (2007) J. Leukoc. Biol. 82:465.

Bioinformatics

• Gene Symbol: ROBO1
• Uniprot:
  • NP_598334.1()