ROBO4 Antibody (265721) [Unconjugated]

HUVEC cells were stained with Mouse Anti-Human ROBO4 Monoclonal Antibody (Catalog # MAB24541, filled histogram) or isotype control antibody (Catalog # MAB004, open histogram) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

• Reactivity: Hu
• Applications: WB, Flow, CyTOF-ready
• Clone: 265721
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

ROBO4 Antibody (265721) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human ROBO4
  Gln28-Arg467
  Accession # Q8WZ75
• Specificity: Detects human ROBO4 in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant rat ROBO1, recombinant human (rh) ROBO2, or rhROBO3 is observed.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: ROBO4
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Western Blot 1 ug/mL

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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

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

Alternate Names for ROBO4 Antibody (265721) [Unconjugated]

• ECSM4
• FLJ20798
• Magic roundabout
• MGC133352
• MGC133353
• MRB
• ROBO4
• roundabout homolog 4
• roundabout homolog 4, magic roundabout (Drosophila)

Background

ROBO4, also called magic roundabout, is a ~150 kDa glycoprotein belonging to the ROBO family (1). ROBOs are molecular guidance receptors that typically interact with Slit ligands to regulate axon guidance and neuronal migration (2). Unlike other family members, ROBO4 is mainly restricted to the vascular endothelium (1, 2). Expression in early hematopoietic progenitors is also reported (3). The human ROBO4 cDNA encodes 1012 amino acids (aa), including a 27 aa signal sequence, a 440 aa extracellular domain (ECD) containing two C2‑type Ig domains and two fibronectin type III (FNIII) domains, a transmembrane domain and an intracellular domain. ROBO4 diverges from other ROBO proteins in the number of Ig, FNIII and cytoplasmic CC domains (1, 4). Within the ECD, human ROBO4 shares 80%, 80%, 87% and 88% aa identity with mouse, rat, bovine and canine ROBO4, respectively. Vascular endothelial ROBO4 is expressed at highest levels in during development and vascular remodeling, including tumor angiogenesis (1, 2, 4‑6). It is proposed to contribute to vascular stability. Consistent with this, endogenous ROBO4 is concentrated in the vascular stalk and sprouts rather than tip cells and appears to protect newly formed blood vessels against VEGF‑induced vascular leak (6‑9). ROBO4 binding of Slit proteins has been variably reported, and when detected may be mediated by ROBO4/ROBO1 heterodimers (2, 4‑7, 9‑13). ROBO4 is also variably reported to stimulate or inhibit cell migration or filopodia formation (2, 4‑13). Effects on cell movement may be mediated through intracellular binding of WASP‑, Ras/Rac/Rho‑, Mena‑, Src‑ or Paxillin‑related proteins, all of which affect the cytoskeleton (5‑7, 10‑12). Recombinant soluble ROBO4 ECD can antagonize endothelial cell migration and in-vivo angiogenesis (13). 

1. Huminiecki, L. et al. (2002) Genomics 79:547.
2. Legg, J.A. et al. (2008) Angiogenesis 11:13.
3. Shibata, F. et al. (2009) Stem Cells 27:183.
4. Park, K.W. et al. (2003) Dev. Biol. 261:251.
5. Seth, P. et al. (2005) Biochem. Biophys. Res. Commun. 332:533.
6. Jones, C.A. et al. (2008) Nat. Med. 14:448.
7. Jones, C.A. et al. (2009) Nat. Cell Biol. 11:1325.
8. Chen, H. et al. (2010) Adv. Exp. Med. Biol. 664:457.
9. London, N.R. et al. (2009) J. Thromb. Haemost. 7:57.
10. Verissimo, A.R. et al. (2009) Biochem. Soc. Trans. 37:1214.
11. Sheldon, H. et al. (2009) FASEB J. 23:513.
12. Kaur, S. et al. (2008) BMC Cell Biol. 9:61.
13. Suchting, S. et al. (2005) FASEB J. 19:121.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ROBO4 Antibody (MAB24541)(1)

Publication using MAB24541

• Guijarro-Munoz I, Cuesta AM, Alvarez-Cienfuegos A, Geng JG, Alvarez-Vallina L, Sanz L The axonal repellent Slit2 inhibits pericyte migration: potential implications in angiogenesis. Exp. Cell Res., 2011-12-14;318(4):371-8. 2011-12-14 [PMID: 22198087] (Flow Cytometry, Human)

Bioinformatics

• Gene Symbol: ROBO4
• Uniprot:
  • Q8WZ75()