Recombinant Human ROBO4 Fc Chimera Protein, CF

Catalog #: 2366-RB Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
2366-RB-050

R&D Systems Recombinant Proteins and Enzymes

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Recombinant Human ROBO4 Fc Chimera Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain

Endotoxin Level

<0.1 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its binding ability in a functional ELISA. When recombinant human ROBO4 Fc Chimera is immobilized at 5 μg/mL, biotinylated recombinant rat UNC5H2/Fc Chimera binds with an apparent K_D< 20 nM.

Source

Mouse myeloma cell line, NS0-derived human ROBO4 protein

Human ROBO4 (Gln28 - Arg467) Accession # Q8WZ75	IEGRMD	Human IgG₁ (Pro100 - Lys330)
N-terminus		C-terminus

Accession #

Q8WZ75

N-terminal Sequence
Analysis

No results obtained: Gln28 predicted

Structure / Form

Disulfide-linked homodimer

Predicted Molecular Mass

74 kDa (monomer)

SDS-PAGE

90-100 kDa, reducing conditions

Product Datasheets

2366-RB

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

2366-RB

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 400 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it 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. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Reconstitution Calculator

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). R&D Systems quality control testing of the ROBO4 ECD has demonstrated high‑affinity binding of the Netrin receptor UNC5H2 (UNC5B).

References