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Human ROBO4 Antibody

Catalog #: MAB24541
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MAB24541
MAB24541-SP
Detection of ROBO4 in HUVEC Human Cells by Flow Cytometry.
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Human ROBO4 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 IgG2B Clone # 265721
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human ROBO4
Gln28-Arg467
Accession # Q8WZ75
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
Human ROBO4
Flow Cytometry
2.5 µg/106 cells
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 

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.

Scientific Data

Flow Cytometry Detection of ROBO4 antibody in HUVEC Human Cells antibody by Flow Cytometry. View Larger

Detection of ROBO4 in HUVEC Human Cells by Flow Cytometry. Human umbilical vein endothelial cells were stained with Mouse Anti-Human ROBO4 Monoclonal Antibody (Catalog # MAB24541, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG F(ab')2Secondary Antibody (Catalog # F0101B).

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

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 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). 

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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Citation for Human ROBO4 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. The axonal repellent Slit2 inhibits pericyte migration: potential implications in angiogenesis.
    Authors: Guijarro-Munoz I, Cuesta AM, Alvarez-Cienfuegos A, Geng JG, Alvarez-Vallina L, Sanz L
    Exp. Cell Res., 2011-12-14;318(4):371-8.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry

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