Human ROBO4 Alexa Fluor® 350-conjugated Antibody

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FAB2454U-100UG
R&D Systems Antibodies
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Human ROBO4 Alexa Fluor® 350-conjugated Antibody Summary

Species Reactivity
Human
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
Monoclonal Mouse IgG2a Clone # 265703
Immunogen
Mouse myeloma cell line NS0-derived recombinant human ROBO4
Gln28-Arg467
Accession # Q8WZ75
Formulation
Supplied 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.
Label
Alexa Fluor 350 (Excitation= 346 nm, Emission= 442 nm)

Applications

Recommended Concentration
Sample
Flow Cytometry
0.25-1 µg/106 cells
HUVEC human umbilical vein endothelial cells

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.

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: ROBO4

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 ROBOs 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). 

References
  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.
Long Name
Roundabout Homolog 4
Entrez Gene IDs
54538 (Human); 74144 (Mouse)
Alternate Names
ECSM4; FLJ20798; Magic roundabout; MGC133352; MGC133353; MRB; ROBO4; roundabout homolog 4; roundabout homolog 4, magic roundabout (Drosophila)

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