Frizzled 2, Fc Chimera, Recombinant, Mouse (FZD2)

Wnt signaling is involved in variety of developmental processes including cell fate determination, cell polarity, tissue patterning and control of cell proliferation. Members of the Frizzled family of proteins serve as receptors for the Wnt signaling pathway. The founding member of this family was identified in Drosophila based on its role in tissue polarity in the adult cuticle and named for the disorganized appearance of bristle hairs on the mutant. Ten mammalian Frizzled genes have been identified to date. The predicted structure of Frizzled proteins is similar among all family members, containing a divergent N-terminal signal peptide, a highly conserved extracellular cysteine-rich domain, a variable-length linker region, a seven-pass transmembrane domain, and a variable-length C-terminal tail. One of the most conserved regions of the Frizzled proteins is the extracellular cysteine-rich domain (CRD) which spans approximately 120aa and contains 10 invariant cysteines (1). Mouse Frizzled-2 shows 100% amino acid identity to human and rat Frizzled-2 in the CRD region. Frizzled-2 expression is greater in embryonic than adult tissues, with heart, brain, lung, kidney and gut showing the highest levels (2). In addition, Frizzled-2 is expressed during migration and proliferation of neural crest cells that populate the heart and great arteries (3). However, Frizzled-2 may not be strictly involved in developmental processes,\nas its expression is also upregulated in myofibroblasts during tissue repair after myocardial infarction (4). Frizzled-2 is implicated in the Wnt/Ca2+ pathway, a mechanism by which Wnt-5a signaling results in calcium release from intracellular stores and activation of PKC and calmodulin-dependent protein kinase (5). Secreted frizzled related proteins (sFRPs), antagonists of the Wnt pathway, have been shown to interact with both Wnt ligands and Frizzled receptors (6).\n\nSource: \nN- Human CD33 Signal Peptide (Met1-Ala16)- Mouse Frizzled-2 (Gln29-Leu168)- IEGRMD- Human IgG1 (Pro100-Lys330)- C\n\nA DNA sequence encoding the signal peptide from human CD33 joined with the extracellular\nCysteine Rich Domain (CRD) of mouse Frizzled-2, amino acid residues 29-168 (Malik, T.H. and\nR.A. Shivdasani, 2000, J. Biochem. 349: 829-834) was fused to the Fc region of human IgG1 via a peptide linker. The chimeric protein was expressed in a mouse myeloma cell line, NS0.\n\nMolecular Mass: \nThe recombinant mature mouse Frizzled-2/Fc chimera, generated by proteolytic removal of the\nsignal peptide, is a disulfide-linked homodimeric protein. The N-terminal amino acid sequence of\nrecombinant mature Frizzled-2/Fc was blocked, consistent with having Gln29 as the first residue.\nEach subunit of Frizzled-2/Fc chimera has a calculated molecular mass of 42.3kD. As a result of\nglycosylation, the recombinant protein migrates as a 50-55kD protein in SDS-PAGE under reducing conditions.\n\nEndotoxin Level: \n(same/less than) 1.0 EU per 1ug of the cytokine as determined by the LAL method.\n\nActivity: \nMeasured by its ability to bind sFRP-2 in a functional ELISA. Immobilized mFrizzled-2/Fc at 2ug/ml (100ul/well) will bind rmsFRP-2 with a linear range of 1.6 -100ng/ml in a functional ELISA.\n\nReconstitution: \nIt is recommended that sterile PBS be added to the vial to prepare a working stock solution of no less than 100ug/ml. The carrier-free protein should be used immediately upon reconstitution to avoid losses in activity due to non-specific binding to the inside surface of the vial. For long term storage as a dilute solution, a carrier protein (e.g. 0.1% HSA or BSA) should be added to the vial.\n\nStorage:\nLyophilized samples are stable for up to twelve months from date of receipt at -20 degrees C to -70 degrees C. Upon reconstitution, this cytokine, in the presence of a carrier protein, can be stored under sterile conditions at 2 degrees to 8 degrees C for 2 weeks or at -20 degrees C to -70 degrees C in a manual defrost freezer for three months without detectable loss of activity. Avoid repeated freeze-thaw cycles.

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SPECIFICATIONS

Catalog Number

F6401-09

Size

50ug

Form

Supplied as a lyophilized powder in 90mM Citrate, 0.26M sodium chloride, 0.25M Tris, pH 7.3.

Purity

(same/more than) 90%, as determined by SDS-PAGE and visualized by silver stain.

References

1. Wang, Y. et al., 1996, J. Biol. Chem 271:4468. 2. Malik, T. and R. Shivdasani, 2000, Biochem. J. 349:829. 3. van Gijn, M. et al., 2001, Anat. Embryol. 203:185. 4. Toyofuku, T. et al., 2000, J. Cell Biol. 150:225. 5. Kuhl, M. et al., 2000, Trends Genet. 16: 279. 6. Bafico, A. et al., 1999, J. Biol. Chem. 274:16180.