hh Human ROBO4 Antibody MAB24541: R&D Systems

Human ROBO4 Antibody

(1 citations)   
  • 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 as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
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.
Data Examples
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')2 Secondary Antibody (Catalog # F0101B).
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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)
Related Research Areas
Citations:

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 Citations: 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., 2012;318(4):371-8.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG2B Isotype Control

Ctrl MAB004 32  
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Mouse IgG2B Isotype Control

Ctrl MAB0041 7  
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Mouse IgG2B Isotype Control

Ctrl MAB0042 6
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Secondary Antibodies
Description Application Cat# Citations Images  

Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Mouse IgG Antibody

WB AF007 4
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rat Anti-Mouse IgG2B APC-conjugated Antibody

Flow F0133 1  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Rat Anti-Mouse IgG2B Fluorescein-conjugated Antibody

Flow F0134 1
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Rat Anti-Mouse IgG2B PE-conjugated Antibody

Flow F0132  
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Mouse IgG2B Antibody

Flow MAB0043  
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