• Purity

  >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie? Blue Staining.

• Endotoxin Level

  <0.01 EU per 1 μg of the protein by the LAL method.  

• Activity

  Measured by its binding ability in a functional ELISA. When Recombinant Rat UNC5H2 Fc Chimera (Catalog # ) is immobilized at 5 μg/mL, Recombinant Human Netrin-1 binds with an apparent K    _d <1 nM. Measured in a cell proliferation assay using RT4?D6P2T rat schwannoma cells. The ED    ₅₀ for this effect is 0.5-2.0 μg/mL.

• Source

  Mouse myeloma cell line, NS0-derived

  Human Netrin-1 (Val22-Ala604) Accession # O95631	HPGGGSGGGSGGGS	6-His tag
  N-terminus		C-terminus

• Accession #

• N-terminal Sequence    
  Analysis

  Val22

• Predicted Molecular Mass

  67.5 kDa

• SDS-PAGE

  80-85 kda, reducing conditions

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.

Background: Netrin-1

Human Netrin-1 (netr: Sanskrit for “one who guides”) is a 75 kDa glycoprotein that is closely related to the laminin  gamma  domain and functions as a chemoattractive or chemorepulsive guidance cue in the central nervous system (CNS) during development (1, 2). The protein is synthesized as a 604 amino acid (aa) precursor that contains a 24 aa signal sequence and a 580 aa mature chain. Residues 46-283 constitute a laminin N-terminal domain (domain VI), while 285-453 make up three laminin-type epidermal growth factor?like domains. There is a final domain that runs from aa 487-601 that qualifies as a Netrin-1 like domain. It is also known as domain C in the context of C. elegans. There are four potential sites for N?linked glycosylation. Human Netrin-1 is 99% aa identical to mouse and rat Netrin-1. Netrin-1 is expressed in adult and embryonic tissues. In the adult, the protein is expressed in Schwann cells, oligodendrocytes and multiple neurons. In embryonic tissues, Netrin-1 is found in somatic mesoderm, heart, branchial pouch, and neuroepithelium. Netrin-1 is a secreted protein that, in addition to its involvement in outgrowth and migration orientation in the developing CNS, plays a significant role in the morphogenesis of endothelial cells and vascular smooth-muscle cells. It is also involved in the processes of cytoskeleton reorganization, angiogenesis, epithelial cell adhesion, and cell migration in the lungs, mammary gland, and pancreas (1, 3).

Netrin-1 effects are controlled through different transmembrane receptors (2). Four of the receptors exist in the Unc-5 (Unc=uncontrolled behaviorally) family of proteins, and these include Unc5h1, Unc5h2, Unc5h3/RCM, and Unc5h4. There are also two receptors that belong to the UNC-40 family of molecules. The first is DCC (deleted in colorectal cancer), and the second is neogenin (newly?generated). UNC-5 receptors are noted to mediate the attraction response of axons and netrins. However, UNC-5 molecules, when co?expressed with DCC in the presence of Netrin-1, will mediate repulsion by varying cellular calcium levels (3). The adenosine A2b receptor may also be involved in chemoattraction, either by binding directly with Netrin-1 or by serving as a co?receptor for DCC. These receptors are known as dependence receptors because they depend on their ligand, in this case Netrin-1, for survival (4). Unbound, the receptors induce a specific death signal (4). It is the dysregulation of these receptor systems that may have important roles in tumor biology. DCC and UNC5 proteins make up a system for either initiating or inhibiting apoptosis (4), and it is now believed that Netrin-1 and its dependence receptors play a major role in tumor biology (4-6).

• References:

1. Dakouane-Giudicelli, M. et al. (2010) J. Histochem. Cytochem. 58:73.

2. Masuda, T. et al. (2008) J. of Neurosci. 28:10380.

3. Bernet, A. and P. Mehlen (2007) Bull. Cancer 94:E12.

4. Cirulli, V. and M. Yebra, (2007) Nat. Rev. Mol. Cell. Biol. 8:296.

5. Dumartin, L. et al. (2010) Gastroenterology 137:1595.

6. Mazelin, L. et al. (2004) Nature 431:80.

• Entrez Gene IDs:

  9423 (Human); 18208 (Mouse); 396389 (Chicken)

• Alternate Names:

  netrin 1; Netrin1; Netrin-1; NTN1; NTN1Lnetrin 1, mouse, homolog of