Application

Coating a plate well (6 well plate) with this recombinant CDH13 protein in a neuronal cell specific medium at 1-10 mg / well allows for 1) human neuronal cell and vascular endothelial cell / receptor interaction studies or 2) human neuronal and vascular endothelial cell/receptor interaction studies in vitro.Use this procedure as a guideline to determine optimal coating conditions for the culture system of choice.1. Thaw CDH13 and dilute to desired concentration using serum-free medium or PBS. The final solution should be sufficiently dilute so that the volume added covers the surface evenly (1-10 µg/well, 6 well plate).2. Add appropriate amount of diluted material to culture surface. 3. Incubate at room temperature for approximately 1.5 hours. 4. Aspirate remaining material. 5. Rinse plates carefully with water and avoid scratching bottom surface of plates. 6. Plates are ready for use. They may also be stored at 2-8 °C damp or air dried if sterility is maintained.

Preparation Note

The full-length extracellular domain of the human CDH13 gene (139-693 aa) was constructed with 31 N-terminal T7/HIS-tag and expressed in E. coli as inclusion bodies. The final product was refolded using our unique “temperature shift inclusion body refolding” technology and chromatographically purified.

Sequence

MASMTGGQQMGRGHHHHHHGNLYFQGGEFELSIVVSPILIPENQRQPFPRDVGKVVDSDRPERSKFRLTGKGVDQEPKGIFRINENTGSVSVTRTLDREVIAVYQLFVETTDVNGKTLEGPVPLEVIVIDQNDNRPIFREGPYIGHVMEGSPTGTTVMRMTAFDADDPATDNALLRYNIRQQTPDKPSPNMFYIDPEKGDIVTVVSPALLDRETLENPKYELIIEAQDMAGLDVGLTGTATATIMIDDKNDHSPKFTKKEFQATVEEGAVGVIVNLTVEDKDDPTTGAWRAAYTIINGNPGQSFEIHTNPQTNEGMLSVVKPLDYEISAFHTLLIKVENEDPLVPDVSYGPSSTATVHITVLDVNEGPVFYPDPMMVTRQEDLSVGSVLLTVNATDPDSLQHQTIRYSVYKDPAGWLNINPINGTVDTTAVLDRESPFVDNSVYTALFLAIDSGNPPATGTGTLLITLEDVNDNAPFIYPTVAEVCDDAKNLSVVILGASDKDLHPNTDPFKFEIHKQAVPDKVWKISKINNTHALVSLLQNLNKANYNLPIMVTDSGKPPMTNITDLRVQVCSCRNSKVDCNAAG