Recombinant Rat Agrin

Agrin is a heparin sulfate proteoglycan secreted by the nerve, whose best characterized role is in the development of the neuromuscular junction during embryogenesis. Agrin is involved in the aggregation of acetylcholine receptors (AChR) during synaptogenesis (1). This activity has been shown to be mediated via a receptor complex that includes a receptor-like tyrosine kinase specific to the skeletal muscle termed muscle-specific kinase (MuSK) (2). Agrin also binds several other proteins on the surface of muscle, including dystroglycan and laminin, which are additional binding steps that are required to stabilize the neuromuscular junction. Agrin contains a number of structural domains, including regions of homology to laminin, Kazal protease inhibitors, and EGF repeats. In addition, there are several isoforms of Agrin which are produced by alternative splicing at sites in the C-terminal half. These isoforms have been shown to have significant differences in AChR clustering activities (3).

The biological activity of Rat AGR was determined by its ability to induce acetylcholine receptor clustering on myotubes differentiated from C2C12 mouse myoblasts. The expected ED₅₀ 2-6 ng/ml.

Histidine Tag

mhhhhhhaat gttaamatar attvsrlpas svtprvypsh tsrpvgrtta ppttrrpptt atnmdrprtp ghqqpskscd sqpclhggtc qdqdsgkgft csctagrggs vcekvqppsm pafkghsfla fptlrayhtl rlalefrale teglllyngn argkdflala lldgrvqfrf dtgsgpavlt slvpvepgrw hrlelsrhwr qgtlsvdget pvvgespsgt dglnldtnly vggipeeqva mvldrtsvgv glkgcirmld innqqlelsd wqraavqssg vgecgdhpcl pnpchggalc qaleagmflc qcppgrfgpt cadekspcqp npchgaapcr vlssggakce cplgrsgtfc qtvletagsr pfladfngfs ylelkglhtf erdlgekmal emvflargps glllyngqkt dgkgdfvsla lhnrhlefcy dlgkgaavir skepialgtw vrvflerngr kgalqvgdgp rvlgespksr kvphtmlnlk eplyiggapd fsklargaav ssgfsgviql vslrghqllt qehvlravdv spfadhpctq algnpclngg scvpreatye clcpggfsgl hcekglveks vgdletlafd grtyieylna vieseltnei paekalqsnh felslrteat qglvlwigka aeradymala ivdghlqlsy dlgsqpvvlr stvkvntnrw lrirahrehr egslqvgnea pvtgssplga tqldtdgalw lgglqklpvg qalpkaygtg fvgclrdvvv ghrqlhlled avtkpelrpc ptp (Pro1788-Ser1798 deleted)

Met

Lyophilized

The predicted molecular weight of Recombinant Rat AGR is Mr 90 kDa. However, the actual molecular weight as observed by migration on SDS-PAGE is Mr 100 kDa.

90

This recombinant protein was 0.2 µm filtered and lyophilized from modified Dulbecco’s phosphate buffered saline (1X PBS) pH 7.2 – 7.3 with no calcium, magnesium, or preservatives.

This lyophilized protein is stable for six to twelve months when stored desiccated at -20°C to -70°C. After aseptic reconstitution, this protein may be stored at 2°C to 8°C for one month or at -20°C to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. See Product Insert for exact lot specific storage instructions.

Next Day Ambient

1. McMahan, UJ. et al. (1987) Prog. Brain Res. 71:391
2. Yancopoulos, GD. et al. (1996) Cell 85:513
3. McMahan, UJ. et al. (1992) Current Op. Cell Biol. 4:869