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Recombinant Human ROBO3 Fc Chimera Protein, CF

Bio-Techne includes R&D Systems | Catalog # 3076-RB

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3076-RB-050

Key Product Details

Source

NS0

Accession #

Structure / Form

Disulfide-linked homodimer

Conjugate

Unconjugated

Applications

Binding Activity

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human ROBO3 protein
Human ROBO3
(Leu40-Ser545)
Accession # Q96MS0
IEGRMD Human IgG1
(Pro100-Lys330)
N-terminus C-terminus

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<0.01 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Leu40

Predicted Molecular Mass

82 kDa (monomer)

SDS-PAGE

95-120 kDa, reducing conditions

Activity

Measured by its binding ability in a functional ELISA.
Biotinylated recombinant mouse Slit-3 captured on a streptavidin-coated plate at 1 µg/mL can bind Recombinant Human ROBO3 Fc Chimera with an apparent Kd <10 nM.

Formulation, Preparation and Storage

3076-RB
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution
Reconstitute at 100 μg/mL in sterile PBS.

Reconstitution Buffer Available:
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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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: ROBO3

Human ROBO3 is a 200 kDa member of the 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). Human ROBO3 is a type I transmembrane glycoprotein that is synthesized as a 1386 amino acid (aa) precursor. It contains a 20 aa signal sequence, an 871 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 474 aa cytoplasmic region (5, 6). The ECD contains five C2-type Ig-like domains (aa 64-531) and three fibronectin (FN) type III domains (aa 555-863). The cytoplasmic region contains three of four possible 15-20 aa long CC (conserved cytoplasmic) motifs that are found in ROBO-1 (7, 8). Human ROBO3 has multiple isoforms. An alternate start site generates a 1365 aa A isoform and a 1341 aa B isoform. These two mature forms differ only over the first 26 and 2 amino acids of the N-terminus, respectively. There are multiple point mutations and insertions in the ROBO3 gene. Three result in truncated forms. One is 456 aa in length and ends after the Ig-like domain #4. A second is 770 aa in length and ends in the third FN domain. A third isoform is truncated after aa 1108 in the cytoplasmic region after CC2. At least one alternate splice form is also reported. It shows a 10 aa substitution between aa 1025 - 1034, followed by truncation. Human ROBO3 ECD is 84% and 91% aa identical to the ECD in mouse and canine ROBO3, respectively. Normally, axons originating on one side of the spinal cord are inhibited from crossing to the other side by a SLIT2-ROBO1 interaction at the midline. ROBO3 is permissive for this event. It is unclear how this is accomplished. One possibility is that it binds directly to ROBO1, blocking SLIT activation. A second possibility involves ROBO3 binding to SLIT2 in a nonproductive interaction. However, only ROBO3 Form B is known to bind to SLIT2 (9-11).

References

  1. Rajagopalan, S. et al. (2000) Neuron 28:767.
  2. Guthrie, S. (2004) Curr. Biol. 14:R632.
  3. Guthrie, S. (2001) Curr. Biol. 11:R300.
  4. Seeger, M. et al. (1993) Neuron 10:409.
  5. Jen, J.C. et al. (2004) Science 304:1509.
  6. Swiss-Prot. Accession # Q96MS0.
  7. Bashaw, G.J. et al. (2000) Cell 101:703.
  8. Kidd, T. et al. (1998) Cell 92:205.
  9. Camurri, L. et al. (2005) Mol. Cell. Neurosci. 30:485.
  10. Sabatier, C. et al. (2004) Cell 117:157.
  11. Mambetisaeva, E.T. et al. (2005) Dev. Dyn. 233:41.

Long Name

Roundabout Homolog 3

Alternate Names

RIG1

Entrez Gene IDs

64221 (Human); 19649 (Mouse)

Gene Symbol

ROBO3

UniProt

Additional ROBO3 Products

Product Documents for Recombinant Human ROBO3 Fc Chimera Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human ROBO3 Fc Chimera Protein, CF

For research use only

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