Recombinant Human LRRN3/NLRR-3 Catalog Number: 5398-LR

DESCRIPTION Source Mouse myeloma cell line, NS0­derived Val23­Thr628, with a C­terminal 6­His tag Accession # AAH35133

N­terminal Sequence Val23 Analysis Predicted Molecular 69.3 kDa Mass

SPECIFICATIONS SDS­PAGE 90­95 kDa, reducing conditions Activity Measured by the ability of the immobilized protein to enhance the adhesion of Saos­2 human osteosarcoma cells to human Fibronectin. When 3 x 104 cells per well are added to rhNLRR­3 coated plates (5 μg/mL, 100 μL/well), approximately 50­70% will adhere after 1 hour at 37 ° C.

Endotoxin Level <0.10 EU per 1 μg of the protein by the LAL method.

Purity >90%, by SDS­PAGE under reducing conditions and visualized by silver stain. Formulation Lyophilized from a 0.2 μm filtered solution in PBS. See Certificate of Analysis for details.

PREPARATION AND STORAGE Reconstitution Reconstitute at 200 μg/mL in sterile 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 freeze­thaw cycles. l 12 months from date of receipt, ­20 to ­70 °C as supplied. l 1 month, 2 to 8 °C under sterile conditions after reconstitution. l 3 months, ­20 to ­70 °C under sterile conditions after reconstitution.

BACKGROUND Neuronal leucine­rich repeat 3 (NLRR­3) is an 85 kDa, N­glycosylated member of a CNS­associated family of transmembrane molecules that includes AMIGO, FLRT, LINGO, LRIG, SLITRK, and Trk subgroups (1, 2). Mature human NLRR­3 consists of a 606 amino acid (aa) extracellular domain (ECD) with 13 LRRs, an RGD integrin­ binding motif, one Ig­like domain, and one fibronectin type III­like domain, a 21 aa transmembrane segment, and a 59 aa cytoplasmic domain (3). Within the ECD, human NLRR­3 shares 91% aa sequence identity with mouse and rat NLRR­3. NLRR­3 is expressed in specific areas of the developing and adult CNS, and more weakly in other tissues including lung, liver, kidney, and adrenal gland (2 ­ 5). It is preferentially expressed in the third layer of the cerebral cortex in contrast to other NLRRs which are more diffusely distributed between cortical layers (6). During mouse development, its expression pattern is generally complementary to that of SLITRK6 in select brain regions, plus the cochlea, eye, tongue, teeth, and nasal mesenchyme (7). NLRR­3 is upregulated in the cerebral cortex following injury (3, 6). Its expression is dependent on NGF stimulation and is strongest in NGF responsive TrkA+ neuroblastoma cell lines that are weakly aggressive (3). The cytoplasmic domain of NLRR­3 contains two Yxxf motifs which are required for its clathrin­mediated internalization (8). The association of NLRR­3 with clathrin and β­adaptin promotes the enhancement and prolongation of EGF induced signaling (8).

References: 1. Chen, Y. et al. (2006) Brain Res. Rev. 51:265. 2. Haines, B.H. et al. (2005) Dev. Biol. 281:145. 3. Hamano, S. et al. (2004) Int. J. Oncol. 24:1457. 4. Taniguchi, H. et al. (1996) Brain Res. Mol. Brain Res. 36:45. 5. Fukamachi, K. et al. (2001) Biochem. Biophys. Res. Commun. 287:257. 6. Ishii, N. et al. (1996) Brain Res. Mol. Brain Res. 40:148. 7. Aruga, J. (2003) Gene Expr. Patterns 3:727. 8. Fukamachi, K. et al. (2002) J. Biol. Chem. 277:43549.

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