Immunoglobulin Superfamily Member 4B, Recombinant, Human (IGSF4B, Nectin-Like 1, Necl-1, TSLL1, CADM3, SynCAM-3)

Human IGSF4B (immunoglobulin superfamily member 4B), also called CADM3, TSLL1, Necl-1 and SynCAM-3, is a neural tissue-specific member of the nectin-like family of immunoglobulin superfamily molecules. It is a 48-50kD type I transmembrane (TM) glycoprotein \nthat is concentrated at non-junctional contact sites of neuronal axons and glial processes. \nIn myelinated peripheral nerve fibers, IGSF4B is concentrated at nodes of Ranvier in contact sites of Schwann cells. The 398 amino acid (aa) human IGSF4B contains a 24 aa signal sequence, a 306 aa extracellular domain (ECD), a 21 aa transmembrane domain and a 47 aa cytoplasmic domain. The ECD contains an N-terminal V-type Ig-like domain that is responsible for Ca ++ \n-independent homophilic and heterophilic interactions with Nectin-1, Nectin-3 or Necl-2 (IGSF-4) in trans. It also contains two C2-type Ig-like domains that are responsible for Ca ++ -independent homophilic dimerization in cis that is thought to precede trans interaction. \n The cytoplasmic domain binds members of the MAGUK guanylate kinase subfamily, such as Dlg3, Pals2 and CASK. These activities are thought to play roles in adhesion and architecture at \nthe synapse. \n\nWe have also shown that IGSF4B promotes in vitro outgrowth of cortical neurons. Of two known splice variants, one shows an insertion of 34 aa near the mature N-terminus and probably represents a 60kD form. A smaller variant shows deletions of aa # 185-231, and 281-398, which includes the first C2-type Ig-like domain and the TM segment. The IGSF4B ECD is highly conserved, sharing 95-96% aa identity between human mouse, rat, dog and cow sequences. The ECD of IGSF-4A, B, C and D proteins share 35-50% aa identity. \n \nMolecular Mass:\nBased on N-terminal amino acid sequencing, the recombinant human IGSF4B protein starts at Pro 21 and has a predicted molecular mass of approximately 34.5kD. As a result of glycosylation, the recombinant monomer migrates as an approximately 38-41kD protein on a SDS-PAGE gel. The preparation may contain human IGSF4B that starts at Asn 25. \n \nEndotoxin Level: < 1.0 EU per 1ug of the protein as determined by the LAL method. \n\nBiological Activity: Measured by its ability to enhance neurite outgrowth of cortical neurons from E18 rat embryos. \n \nRecommended Dilutions: \nImmobilized rh IGSF4B (at a 3il droplet containing 50 ng) on a nitrocellulose coated microplate is sufficient to significantly enhance neurite outgrowth. \nOptimal dilutions to be determined by the researcher. \n\nHybridoma: \nA DNA sequence encoding the signal peptide from human CD33 joined with the extracellular domain of human IGSF4B (Pro 21-Tyr 329; Accession # Q8N126) was fused with a 6x histidine tag at the C-terminus. The chimeric protein was expressed in a mouse myeloma cell line, NS0. \n\nStorage and Stability:\n\nLyophilized powder may be stored at -20 degrees C. Reconstitute to nominal volume by adding sterile PBS. Aliquot and store at -20 degrees C. Reconstituted product is stable for 1 month at 4 degrees C and 3 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

I7502-B71

Size

50ug

Reactivities

Hum

Form

Supplied as a lyophilized powder in PBS. Reconstitute with sterile PBS to a concentration of >100ug/ml . For long term storage a carrier protein (e.g. 0.1% HSA or BSA) should be added.

Purity

> 90%, as determined by SDS-PAGE and visualized by silver stain. Endotoxin: < 1.0 EU per 1ug of the protein as determined by the LAL method.

References

1. Fukuhara, H. et al., 2001, Oncogene 20:5401. \n2. Kakunaga, S. et al., 2005, J. Cell Sci. 118:1267. \n3. Fukami, T. et al., 2003, Gene 323:11. \n4. Shingai, T. et al., 2003, J. Biol. Chem. 278:35421-35427. \n5. Dong, X. et al., 2006, J. Biol. Chem. 281:10610.

Additional Info

Recognizes human IGSF4B.The IGSF4B ECD is highly conserved, sharing 95-96% aa identity between human mouse, rat, dog and cow sequences. The ECD of IGSF-4A, B, C and D proteins share 35-50% aa identity.