Recombinant Human ROBO3 Fc Chimera Protein, CF

Catalog # Availability Size / Price Qty
3076-RB-050
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human ROBO3 Fc Chimera Protein, CF Summary

Product Specifications

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.
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.
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
Accession #
N-terminal Sequence
Analysis
Leu40
Structure / Form
Disulfide-linked homodimer
Predicted Molecular Mass
82 kDa (monomer)
SDS-PAGE
95-120 kDa, reducing conditions

Product Datasheets

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3076-RB

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

3076-RB

Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS.
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.
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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
Entrez Gene IDs
64221 (Human); 19649 (Mouse)
Alternate Names
FLJ21044; HGPPS; HGPS; RBIG1horizontal gaze palsy with progressive scoliosis; retinoblastoma inhibiting gene 1; RIG1; ROBO3; roundabout (axon guidance receptor, Drosophila) homolog 3; roundabout homolog 3; roundabout, axon guidance receptor, homolog 3 (Drosophila); Roundabout-like protein 3

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