Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Flow Cytometry, CyTOF-ready

Cited:

Flow Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 265721
View all ROBO4 Primary Antibodies »

Product Specifications

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human ROBO4 Antibody

Detection of ROBO4 in HUVEC cells by Flow Cytometry

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.

Applications for Human ROBO4 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: HUVEC human umbilical vein endothelial cells

Western Blot

1 µg/mL
Sample: Human ROBO4

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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.

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.

Calculators

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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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

Alternate Names

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

Entrez Gene IDs

54538 (Human); 74144 (Mouse)

Gene Symbol

ROBO4

UniProt

Q8WZ75

Additional ROBO4 Products

Product Documents for Human ROBO4 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human ROBO4 Antibody

For research use only

Citations for Human ROBO4 Antibody

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